WO2023200875A1 - Octapeptide pour application topique - Google Patents

Octapeptide pour application topique Download PDF

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Publication number
WO2023200875A1
WO2023200875A1 PCT/US2023/018348 US2023018348W WO2023200875A1 WO 2023200875 A1 WO2023200875 A1 WO 2023200875A1 US 2023018348 W US2023018348 W US 2023018348W WO 2023200875 A1 WO2023200875 A1 WO 2023200875A1
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Prior art keywords
composition
skin
seq
expression
octapeptide
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PCT/US2023/018348
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English (en)
Inventor
Alan David Widgerow
John A. Garruto
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ALASTIN Skincare, Inc.
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Publication of WO2023200875A1 publication Critical patent/WO2023200875A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/65—Collagen; Gelatin; Keratin; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations

Definitions

  • Hyaluronic acid plays an important role in cellular and extracellular matrix (ECM) homeostasis in skin.
  • ECM extracellular matrix
  • the HA-bound water in both the dermis and the epidermis is critical for skin hydration. While HA decreases with aging, HA content in skin in fact remains constant with age. The difference is that the HA becomes increasingly tissue-bound and more resistant to extraction.
  • This HA encased within tissue proteins may be limited in its hydrating capacity and lower molecular weight (MW) HA present in these tissue-bound forms may be pro-inflammatory.
  • MW molecular weight
  • Elastin is a key extracellular matrix (ECM) protein that provides stretch, recoil, resilience, and elasticity to tissues and organs, including skin.
  • ECM extracellular matrix
  • Normal levels of elastic fiber production, organization, and integration with other cutaneous extracellular matrix proteins, proteoglycans, and glycosaminoglycans, are key to maintaining healthy skin structure, function, and youthful appearance.
  • Elastin production decreases with age, and elastin degrades upon exposure to environmental insults, particularly sun exposure, causing loss of skin structural integrity and contributing to sagging skin or crepiness and related undesirable changes in appearance.
  • topical compositions which can enhance production of elastic fibers would be highly advantageous for improving and/or restoring appearance, texture, resiliency, and wound-healing capabilities to the skin.
  • Skin aging is associated with a loss of skin moisture and reduced barrier function.
  • Aged or damaged skin can present with various signs of inflammation, which may be visible as increased skin redness, increased visibility of fine blood vessels, and certain skin conditions including, but not limited to, rosacea.
  • Environmental influences, such as excessive UV light exposure may contribute to the visible, progressive changes seen in aged skin.
  • Topical compositions and methods that promote intrinsic high MW HA production or increase elastin production in skin may be helpful to treat various skin conditions.
  • compositions and methods comprising a topical formulation and application of an octapeptide.
  • the compositions and methods described herein reduce skin inflammation, appearance of solar elastosis, and/or signs of skin aging.
  • compositions and methods described herein improve skin health, appearance, and hydration.
  • the compositions and methods described herein increase HA production and/or incraese elastin production.
  • the present disclosure relates to a topical composition for stimulating synthesis of high-molecular- weight hyaluronic acid (high MW HA), the composition comprising a synthetic, isolated, or recombinant polypeptide comprising an amino acid sequence selected from: GPHGVREA (SEQ ID NO: 1), GDGDGASA (SEQ ID NO: 2), GPMGPSGP (SEQ ID NO: 3), GLGPGARA (SEQ ID NO: 4), GPQGFQGP (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), or GPGKNGDD (SEQ ID NO: 7), wherein the polypeptide comprises no more than 20 amino acids.
  • the amino acid sequence is GPHGVREA (SEQ ID NO: 1).
  • the polypeptide is an octapeptide.
  • the polypeptide comprises octapeptide-45.
  • the composition comprises a high molecular weight (MW) hyaluronic acid (HA) or derivatives thereof, wherein the high MW HA has a molecular weight of at least 0.5 MDa. In some embodiments, the high MW HA or derivatives thereof has a molecular weight of about 1 MDa to 4 MDa. In some embodiments, the high MW HA or derivatives comprises sodium hyaluronate. In some embodiments, the sodium hyaluronate comprise a sodium hyaluronate crosspolymer.
  • the composition comprises a synthetic tripeptide.
  • the synthetic tripeptide is tetradecyl aminobutyroylvalylaminobutyric urea tri fluoroacetate.
  • composition comprises a hexapeptide.
  • the hexapeptide is hexapeptide-11.
  • the hexapeptide- 11 is encapsulated in a liposome.
  • the composition comprises lactoferrin.
  • the lactoferrin is encapsulated in a liposome.
  • the composition comprises phosphatidylserine. In some embodiments, the composition comprises Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyl decanone. In some embodiments, the composition comprises a high molecular weight hyaluronic acid, synthetic tripeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyl decanone, or combinations thereof. In some embodiments, the composition comprises hexapeptide-12. In some embodiments, the composition comprises tripeptide-1. In some embodiments, the composition comprises tripeptide-1 and hexapeptide-12.
  • the composition is aqueous.
  • composition is effective to achieve a reduction in inflammation.
  • the reduction in inflammation is assessed by a reduction or a lack of increase in an expression level of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL- 12b), or cluster of differentiation-80 (CD80), or combinations thereof.
  • NOS2 nitric oxide synthase 2
  • TNF tumor necrosis factor
  • IL- 12b interleukin 12
  • CD80 cluster of differentiation-80
  • the composition increases CD44 expression.
  • the composition reduces redness of a skin of an individual after applying the composition.
  • the redness is measured by photography.
  • the composition increases skin hydration in a skin of an individual after applying the composition. In some embodiments, the composition reduces or reverses signs of solar elastosis.
  • the composition upregulates hyaluronic acid synthase (HAS) expression.
  • HAS hyaluronic acid synthase
  • the HAS comprises HAS2.
  • the composition downregulates hyaluronidase expression.
  • the hyaluronidase comprises hyaluronidase 2 (HYAL2).
  • the present disclosure relates to a method for reducing inflammation in a skin of an individual, the method comprising: applying a topical composition comprising a synthetic, isolated, or recombinant polypeptide comprising an amino acid sequence GPHGVREA (SEQ ID NO: 1), GDGDGASA (SEQ ID NO: 2), GPMGPSGP (SEQ ID NO: 3), GLGPGARA (SEQ ID NO: 4), GPQGFQGP (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), or GPGKNGDD (SEQ ID NO: 7), wherein the polypeptide comprises no more than 20 amino acids.
  • the amino acid sequence is GPHGVREA (SEQ ID NO: 1).
  • the polypeptide is an octapeptide.
  • the polypeptide comprises octapeptide-45.
  • the composition comprises a high molecular weight (MW) hyaluronic acid (HA) or derivatives thereof, wherein the high MW HA has a molecular weight of at least 0.5 MDa. In some embodiments, the high MW HA or derivatives thereof has a molecular weight of about 1 MDa to 4 MDa. In some embodiments, the high MW HA or derivatives comprises sodium hyaluronate. In some embodiments, the sodium hyaluronate comprise a sodium hyaluronate crosspolymer. [0024] In some embodiments, the composition comprises a synthetic tripeptide. In some embodiments, the synthetic tripeptide is tetradecyl aminobutyroylvalylaminobutyric urea trifluoroacetate.
  • the composition comprises a hexapeptide.
  • the hexapeptide is hexapeptide-11.
  • the hexapeptide- 11 is encapsulated in a liposome.
  • the composition comprises lactoferrin.
  • the lactoferrin is encapsulated in a liposome.
  • the composition comprises phosphatidylserine. In some embodiments, the composition comprises Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyl decanone.
  • the composition comprises a synthetic tripeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyl decanone, or combinations thereof.
  • the composition comprises hexapeptide-12.
  • the composition comprises tripeptide-1.
  • the composition comprises tripeptide-1 and hexapeptide-12.
  • the composition is aqueous.
  • the reduction in inflammation is assessed by a reduction or a lack of increase in an expression level of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL- 12b), or cluster of differentiation-80 (CD80), or combinations thereof.
  • NOS2 nitric oxide synthase 2
  • TNF tumor necrosis factor
  • IL- 12b interleukin 12
  • CD80 cluster of differentiation-80
  • the composition increases CD44 expression.
  • the composition reduces redness of a skin of an individual after applying the composition.
  • the redness is measured by photography.
  • the composition increases skin hydration in a skin of an individual after applying the composition. In some embodiments, the composition promotes HA synthesis.
  • the composition upregulates hyaluronic acid synthase (HAS) expression.
  • HAS hyaluronic acid synthase
  • the HAS comprises HAS2.
  • the composition downregulates hyaluronidase expression.
  • the hyaluronidase comprises hyaluronidase 2 (HYAL2).
  • the composition reduces appearance of a bruise, an aging spot, or a wrinkle.
  • the composition is applied 1, 2, 3, 4, 5, 6, 7, or 8 times a day.
  • the individual is a human.
  • the present disclosure relates to a topical composition for promoting skin repair, the topical composition comprising a synthetic, isolated, or recombinant octapeptide comprising an amino acid sequence Gly - XI - X2 - X3 - X4 - X5 - X6 - X7.
  • XI is Pro, Asp, or Leu.
  • X2 is His, Gly, Met, or Gin.
  • X3 is Gly, Asp, Pro, or Lys.
  • X4 is Vai, Gly, Pro, Phe, or Asn.
  • X5 is Arg, Ala, Ser, Gin, or Gly.
  • X6 is Glu, Ser, Gly, Arg, or Asp.
  • X7 is Ala, Pro, or Asp.
  • the octapeptides comprises an amino acid sequence feature found in collagen or elastin.
  • the amino acid sequence comprises GPHGVREA (SEQ ID NO: 1), GDGDGASA (SEQ ID NO: 2), GPMGPSGP (SEQ ID NO: 3), GLGPGARA (SEQ ID NO: 4), GPQGFQGP (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8).
  • the amino acid sequence is GPHGVREA (SEQ ID NO: 1).
  • the polypeptide comprises octapeptide-45.
  • the composition comprises a high molecular weight (MW) hyaluronic acid (HA) or derivatives thereof, wherein the high MW HA has a molecular weight of at least 0.5 MDa.
  • the high MW HA or derivatives comprises sodium hyaluronate.
  • the sodium hyaluronate comprise a sodium hyaluronate crosspolymer.
  • the composition comprises a synthetic tripeptide.
  • the synthetic tripeptide is tetradecyl aminobutyroylvalylaminobutyric urea tri fluoroacetate.
  • the composition comprises a hexapeptide.
  • the hexapeptide is hexapeptide-11.
  • the hexapeptide- 11 is encapsulated in a liposome.
  • the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome. [0044] In some embodiments, the composition comprises phosphatidylserine. In some embodiments, the composition comprises Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyl decanone.
  • the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyl decanone, or combinations thereof.
  • the composition comprises hexapeptide-12.
  • the composition comprises tripeptide-1.
  • the composition comprises tripeptide-1 and hexapeptide-12.
  • the composition is aqueous.
  • the reduction in inflammation is assessed by a reduction or a lack of increase in an expression level of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL- 12b), or cluster of differentiation-80 (CD80), or combinations thereof.
  • NOS2 nitric oxide synthase 2
  • TNF tumor necrosis factor
  • IL- 12b interleukin 12
  • CD80 cluster of differentiation-80
  • the composition increases CD44 expression.
  • the composition reduces redness of a skin of an individual after applying the composition.
  • the redness is measured by photography.
  • the composition increases skin hydration in a skin of an individual after applying the composition. In some embodiments, the composition promotes HA synthesis. In some embodiments, the composition promotes elastin production.
  • the composition upregulates hyaluronic acid synthase (HAS) expression.
  • HAS comprises HAS2.
  • the composition downregulates hyaluronidase expression.
  • the hyaluronidase comprises hyaluronidase 2 (HYAL2).
  • the composition reduces or reverses a sign of solar elastosis.
  • the composition stimulates synthesis of high MW HA having a MW of at least 0.5 MDa.
  • the present disclosure relates to a topical composition for increasing the production of elastin in mammalian skin
  • a polypeptide comprising an amino acid sequence selected from the group consisting of GPHGVREA (SEQ ID NO:1), GDGDGASA (SEQ ID NO: 2), GPMGPSGP (SEQ ID NO: 3), GLGPGARA (SEQ ID NO: 4), GPQGFQGP (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), or GPGKNGDD (SEQ ID NO: 7), wherein the polypeptide comprises no more than 20 amino acids; tripeptide-1; and hexapeptide-12.
  • the polypeptide is an octapeptide.
  • the amino acid sequence of the polypeptide comprises GPHGVREA (SEQ ID NO: 1).
  • the octapeptide is octapeptide-45.
  • the composition further comprises hexapeptide- 11, tetradecylaminobutyroylvalylaminoburyric urea trifluoroacetate, lactoferrin, phosphatidylserine, Tremella fuciformis extract, hydroxymethoxyphenyl decanone, or any combination thereof.
  • the composition comprises a high molecular weight (MW) hyaluronic acid (HA) or derivatives thereof, wherein the high MW HA has a molecular weight of at least 0.5 MDa
  • the composition comprises hexapeptide-11.
  • the composition comprises lactoferrin.
  • the composition comprises phosphatidylserine.
  • the composition comprises Tremella fuciformis extract.
  • the composition comprises tetradecylaminobutyroylvalylaminoburyric urea trifluoroacetate.
  • the composition comprises hydroxymethoxyphenyl decanone.
  • any one of the hexapeptide-11, tetradecylaminobutyroylvalylaminoburyric urea trifluoroacetate, lactoferrin, phosphatidylserine, Tremella fuciformis extract, hydroxymethoxyphenyl decanone, or any combination thereof, are encapsulated in a liposome.
  • the composition is aqueous.
  • the composition is effective to increase the production of tropoelastin and microfibrillar-associated protein 4 (MFAP4).
  • the composition is effective to upregulate expression of S100A2, LAMA3, MERTK, or ITGB4.
  • the composition upregulates one or more genes related to pathways related to ECM regulation.
  • the present disclosure relates to a method of treating or restoring aging skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of upregulating expression of MFAP4, S100A2, LAMA3, MERTK, or ITGB4 expression in skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of increasing MF AIM expression in skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of increasing S100A2 expression in skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of increasing LAMA3 expression in skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of increasing MERTK expression in skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of increasing ITGB4 expression in skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of upregulating genes related to pathways related to ECM regulation in skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of increasing MFAP4 expression in skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a method of increasing elastin production in human skin, the method comprising: administering to the skin a composition according to any of the embodiments described herein.
  • the present disclosure relates to a composition according to any of the embodiments described herein, for use in treating or restoring aging skin or increasing elastin production in human skin.
  • the composition increases expression of MFAP4, S100A2, LAMA3, MERTK, or ITGB4 in skin.
  • the composition upregulates expression of genes related to pathways related to ECM regulation in skin.
  • the present disclosure relates to a method for promoting wound healing, comprising administering to the skin the topical composition according to any of the embodiments described herein.
  • the present disclosure relates to a method for promoting wound healing or wound closure, comprising administering to the skin the topical composition according to any of the embodiments described herein.
  • the present disclosure relates to a method for treating a cutaneous wound, comprising administering to the skin the topical composition according to any of the embodiments described herein.
  • FIG. 1 illustrates a graph of changes in fold of mRNA gene expression of Hyaluronic Acid Synthase (HAS2) in dermal fibroblasts following treatment with compounds listed below the bars compared with untreated cells.
  • HAS2 Hyaluronic Acid Synthase
  • FIG. 2A illustrates a graph of changes in fold of mRNA gene expression of Hyaluronic Acid Synthase (HAS2) in keratinocytes following treatment with compounds listed below the bars compared with untreated cells.
  • HAS2 Hyaluronic Acid Synthase
  • FIG. 2B illustrates a graph of changes in fold of mRNA gene expression of Hyaluronidase 2 (HYAL2) in keratinocytes following treatment with compounds listed below the bars compared with untreated cells.
  • HYAL2 Hyaluronidase 2
  • FIG. 3 illustrates a graph of changes in fold of mRNA gene expression of Early Growth Response 3 (EGR3) in keratinocytes following treatment with compounds listed below the bars compared with untreated cells.
  • EGR3 Early Growth Response 3
  • FIG. 4 depicts an SDS-PAGE gel with concentrated supernatant from cultured human dermal fibroblasts demonstrating the effects of treatment with various compounds.
  • FIG. 5 depicts an SDS-PAGE gel with concentrated supernatant from cultured human dermal fibroblasts demonstrating the effects of treatment with various compounds.
  • FIG. 6 illustrates a graph of dose response stimulation of high molecular weight HA production by Octapeptide treatment in dermal fibroblasts.
  • FIG. 7 is a line plot demonstrating a progressive increase in the mean hydration levels on clean skin 15 minutes post cleansing of facial skin.
  • FIG. 8 is a bar graph demonstrating a progressive increase in the mean hydration levels on clean skin following cleansing of facial skin, waiting a first 15-minute period post cleansing, applying the treatment with a composition comprising high MW HA and octapeptide and waiting a second 15-minute period post treatment prior to hydration measurement.
  • FIG. 9 is a table aggregating the results of participant assessment and satisfaction surveys following post cleansing the skin at various time points after the initiation of treatment with a composition comprising high MW HA and octapeptide.
  • FIG. 10 is a table aggregating the results of participant assessment surveys of their skin condition beginning at treatment initiation (Baseline) and at various time points after the initiation of treatment.
  • FIG. 11 is a table of participant satisfaction and a statistical analysis of the differences between baseline ratings and the ratings at various time points after the initiation of treatment.
  • FIG. 12 is a chart comparing mean investigator assessment scores for the given criteria of skin condition at baseline and following 8 weeks of treatment.
  • FIG. 13 is a chart comparing mean investigator assessment scores for skin moisture/hydration at baseline and following 8 weeks of treatment.
  • FIG. 14-21 are photographs, analysis and histology sections from subjects at baseline and at various time points following initiation of treatment with a composition comprising high MW HA and octapeptide, SPF 30+, an optional ultra-light moisturizer post Week 4, and regular skin care using a gentle cleanser.
  • FIG. 14A shows photographs taken at baseline and at Week 4 following treatment to demonstrate improvements to the skin in a female subject, age 39.
  • FIG. 14B shows analysis of photographs from FIG. 14A using the VISIA® Skin Analysis System to demonstrate a reduction in red areas of the face following treatment.
  • FIG. 15 shows photographs taken (from left to right) at baseline, at Week 2, at Week 4, and at Week 8 following treatment to demonstrate improvements to progressive effects of skin improvement in a female subject, age 35.
  • FIG. 16A shows close-up photographs taken at baseline and at Week 8 following treatment to demonstrate improvements in pore size, skin tone and skin texture in a female subject, age 35.
  • FIG. 16B shows analysis of photographs from FIG. 15 taken at baseline and at Week 8 following treatment using the VISIA® Skin Analysis System to demonstrate a reduction in red areas of the face.
  • FIG. 16C shows analysis of photographs from FIG. 15 taken at baseline and at Week 8 following treatment using the VISIA® Skin Analysis System to demonstrate a reduction in pore sizes on the face.
  • FIG. 16D shows close-up photographs of the eye and cheek area in a female subject, age 35 from FIG. 15 taken at baseline and at Week 8 following treatment to demonstrate improvements in fine lines, wrinkles, and redness reduction.
  • FIG. 16E shows analysis of photographs of the eye and cheek area in a female subject, age 35 from FIG. 15 taken at baseline and at Week 8 following treatment using the VISIA® Skin Analysis System to demonstrate a reduction wrinkle density, area, and distribution.
  • FIG. 17A shows photographs taken at baseline and at Week 8 following treatment to demonstrate improvements to the skin tone, texture and redness reduction in a female subject, age 38.
  • FIG. 17B shows analysis of photographs from FIG. 17A using the VISIA® Skin Analysis System to demonstrate an improvement to yield a more even skin texture in areas of the face following treatment.
  • FIG. 18A shows photographs taken (from left to right) at baseline, at Week 2, at Week 4, and at Week 8 following treatment to demonstrate improvements of a progressive reduction in skin inflammation, a reduction in skin redness, and a reduction in visibility of fine blood vessels in a male subject, age 48.
  • FIG. 18B shows close-up photographs of the upper left face from the male subject, age 48 from FIG. 18 A taken at baseline and at Week 8 following treatment to demonstrate improvements rosacea and inflammation.
  • FIG. 18C shows close-up photographs of the right cheek from the male subject, age 48 from FIG. 18A taken at baseline and at Week 8 following treatment to demonstrate skin improvements including reduced visibility of fine blood vessels.
  • FIG. 19A shows close-up photographs of right cheek and mouth area of a female subject, age 73 taken at baseline and at Week 2 following treatment to demonstrate improvements in fine lines, deep wrinkles and skin crepiness.
  • FIG. 19B shows photographs of right side of the face of the female subject, age 73 from FIG. 19A taken at baseline and at Week 4 following treatment to demonstrate progressive improvements in fine lines, deep wrinkles and skin crepiness.
  • FIG. 19C shows photographs of right side of the face of the female subject, age 73 from FIG. 19A taken at baseline and at Week 8 following treatment to demonstrate progressive improvements in fine lines, deep wrinkles and skin crepiness.
  • FIG. 20A-C show histological sections of periauricular biopsies of subjects at baseline and at Week 8 following treatment to demonstrate marked improvement in solar elastotic extracellular matrix (ECM). Paired sections from three separate subjects at the given time points are stained with hematoxylin and eosin. Magnification is 200X.
  • FIG. 21A-C show histological sections of periauricular biopsies of subjects at baseline and at Week 8 following treatment to demonstrate marked upregulation of CD44 expression in epidermal and dermal structures in a third experiment.
  • FIG. 21 A has paired sections from a first subject; magnification at 200X.
  • FIG. 21B-C have paired sections from a second subject; magnification at 200X and 400X respectively.
  • FIG. 22 shows a diagram of the chemical structure of octapeptide-45, comprising an amino acid sequence Gly-Pro-His-Gly-Val-Arg-Glu-Ala.
  • FIGS. 23A-23D are bar graphs showing results from in vitro gene expression studies (percent change versus untreated control) for human dermal fibroblasts cultured and treated with tripeptide-1 + hexapeptide- 12 (TriHex), octapeptide-45, and TriHex + octapeptide-45.
  • FIG. 23A shows results for S100A2 expression
  • FIG. 23B shows results for LAMA3 expression
  • FIG. 23C shows results for MERTK expression
  • FIG. 23D shows results for ITGB4 expression. All treatments were 20 pg/mL.
  • FIG. 24 is a schematic illustration of an ex vivo experimental model used for testing the effect on protein production for peptide compositions according to the present disclosure.
  • FIG. 25 is a schematic illustration of a model showing tropoelastin and MFAP4 production in elastin fiber assembly. (Reproduced from P. Bartosz et al., 291 J. Biol. Chem. 1103-14 (Jan. 2016) ( Figure 10).)
  • FIG. 26 is an illustration of a human skin sample cross-section, indicating the location of the epidermis, papillary dermis, and reticular dermis.
  • FIG. 27 shows stained human skin sample cross-sections for untreated ski (left) and skin treated with tripeptide-1 + hexapeptide- 12 (TriHex) (second from left), octapeptide-45 (second from right), and TriHex + octapeptide-45 (right).
  • the blue stain represents DAPI for nuclei and cells.
  • the red stain represents MFAP4.
  • FIG. 28 shows stained human skin sample cross-sections for untreated ski (left) and skin treated with tripeptide-1 + hexapeptide- 12 (TriHex) (second from left), octapeptide-45 (second from right), and TriHex + octapeptide-45 (right).
  • the blue stain represents DAPI for nuclei and cells.
  • the red stain represents tropoelastin (TE).
  • Skin aging is associated with a loss of skin moisture and reduced barrier function.
  • Aged or damaged skin can present with various signs of inflammation, which may be visible as increased skin redness, increased visibility of fine blood vessels, and certain skin conditions including, but not limited to, rosacea.
  • Environmental influences, such as excessive UV light exposure, may contribute to the visible, progressive changes seen in aged skin.
  • compositions and methods comprising a topical formulation and application of an octapeptide.
  • the compositions and methods described herein reduce skin inflammation, appearance of solar elastosis, and/or signs of skin aging.
  • compositions and methods described herein improve skin health, appearance, and hydration.
  • the composition comprising an octapeptide results in increase synthesis of high MW HA as compared to a skin untreated with the composition.
  • the octapeptide in the composition stimulates synthesis of high MW HA.
  • the compositions increase elastin production.
  • a group of active agents that stimulated high MW HA production in both epidermal and dermal cell types were identified.
  • Members of one class of these active agents e.g. octapeptides
  • endogenous signaling pathways may recognize these short peptide sequences that mimic extracellular matrix (ECM) degradation products and upregulate a natural response that produces high MW HA.
  • ECM extracellular matrix
  • compositions and methods by which to use them were created to maximize hydration capacity in both the epidermis and dermis while encouraging regenerative activity within in the ECM.
  • Application of an octapeptide formulation led to quantifiable increases in skin hydration; concurrent decreases in redness, inflammation, and the appearance of fine lines and wrinkles; and a striking reversal of solar elastotic ECM structure to a healthier and denser ECM network.
  • the compositions comprising an octapeptide increase production of elastin in the skin.
  • the compositions comprise octapeptide-45, tripeptide-1, and hexapeptide-12.
  • the compositions upregulate expression of genes related to ECM regulation in the skin.
  • the compositions increase MFAP4 expression.
  • the compositions increase tropoelastin (TE) production.
  • the compositions upregulate SAI 002 expression.
  • the compositions upregulate LAMA3 expression.
  • the compositions upregulate MERTK expression.
  • the compositions upregulate ITGB4 expression.
  • the compositions upregulate gene expression for one or more pathways related to ECM regulation.
  • compositions comprising an octapeptide and methods of preparing and using such compositions.
  • the compositions and methods comprise one or more peptides for inducing new elastogenesis in human skin or stimulating hyaluronic acid (HA) production.
  • the compositions and methods comprise one or more peptides for stimulating production of high molecular weight HA.
  • the octapeptide comprises octapeptide-45.
  • the octapeptide-45 is a synthetic octapeptide.
  • the octapeptide-45 is a synthetic octapeptide having the chemical structure illustrated in FIG. 22.
  • compositions and methods described herein comprise one or more octapeptides.
  • the one or more octapeptides comprises an octapeptide amino acid sequence H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH.
  • the octapeptide amino acid sequence comprises H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH, wherein Xi is Pro, Asp, or Leu.
  • the octapeptide amino acid sequence comprises H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH, wherein X2 is His, Gly, Met, or Gin.
  • the octapeptide amino acid sequence comprises H - Gly - Xi - X2 - X3 - X4 - Xs - Xe - X7 - OH, wherein X3 is Gly, Asp, Pro, or Lys.
  • the octapeptide amino acid sequence comprises H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH, wherein X4 is Vai, Gly, Pro, Phe, or Asn.
  • the octapeptide amino acid sequence comprises H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH, wherein X5 is Arg, Ala, Ser, Gin, or Gly.
  • the octapeptide amino acid sequence comprises H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH, wherein Xe is Glu, Ser, Gly, Arg, or Asp.
  • the octapeptide amino acid sequence comprises H - Gly - X1 - X2 - X3 - X4- X5 - Xe- X7- OH, wherein X7 is Ala, Pro, or Asp.
  • the octapeptide amino acid sequence comprises H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH, wherein Xi is Pro, Asp, or Leu; wherein X2 is His, Gly, Met, or Gin; wherein X3 is Gly, Asp, Pro, or Lys; wherein X4 is Vai, Gly, Pro, Phe, or Asn; wherein X5 is Arg, Ala, Ser, Gin, or Gly; wherein Xe is Glu, Ser, Gly, Arg, or Asp; and wherein X7 is Ala, Pro, or Asp.
  • the octapeptide amino acid sequence comprises H - Gly - Pro - Met - Gly - Pro - X5 - Gly - Pro - OH, wherein X5 is Ser or Arg.
  • the octapeptide amino acid sequence comprises one or more of GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD.
  • the octapeptide is GPHGVREA.
  • the octapeptide has a chemical structure of octapeptide-45.
  • the octapeptide is octapeptide-45.
  • the octapeptide-45 is GPHGVREA.
  • compositions and methods described herein comprise one or more octapeptides.
  • the one or more octapeptides comprise octapeptides with sequence features found in collagen and/or elastin.
  • the octapeptides with sequence features found in collagen and/or elastin comprise an amino acid sequence GXXXXX.
  • the octapeptides with sequence features found in collagen and/or elastin comprise an amino acid sequence H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH, wherein Xi is Pro, Asp, or Leu; wherein X2 is His, Gly, Met, or Gin; wherein X3 is Gly, Asp, Pro, or Lys; wherein X4 is Vai, Gly, Pro, Phe, or Asn; wherein X5 is Arg, Ala, Ser, Gin, or Gly; wherein Xe is Glu, Ser, Gly, Arg, or Asp; and wherein X7 is Ala, Pro, or Asp.
  • the composition comprises an octapeptide, a hexapeptide, or a tripeptide, or a combination thereof.
  • the composition comprises a peptide with an amino acid sequence features found in collagen and/or elastin.
  • the composition comprises a peptide with an amino acid sequence features similar or identical to chemical or enzymatic degradative product of collagen.
  • the composition comprises a peptide with an amino acid sequence features similar or identical to chemical or enzymatic degradative product of elastin.
  • the compositions and methods comprise one or more peptides for inducing new elastogenesis in human skin or stimulating elastin production.
  • compositions and methods comprise one or more peptides for inducing new elastogenesis in human skin or stimulating collagen production. In some embodiments, the compositions and methods comprise one or more peptides for inducing new elastogenesis in human skin or turnover of older, thin collagen fibers with thicker, healthier collagen fibers. In some embodiments, the compositions and methods comprise one or more peptides for inducing new elastogenesis in human skin or stimulating elastin and collagen production. In some embodiments, the one or more elastin-derived peptides is a hexapeptide. In some embodiments, the one or more elastin-derived peptides comprises an amino acid sequence XGVXXG.
  • the one or more elastin-derived peptides comprises a sequence according to amino acid sequence IGVAPG, VGVAPG, or VGVTAG.
  • Elastin-derived peptides e.g., XGVXXG
  • compositions and methods described herein comprise one or more tripeptides, one or more hexapeptides, one or more octapeptides, or combinations thereof.
  • the one or more peptides (e.g., elastin-derived peptide) is synthetic.
  • compositions and methods described herein comprise a mixture of peptides (e.g., an elastin-derived peptide) that provide synergistic effects (e.g., gene expression).
  • the compositions and methods comprising one or more peptides with amino acid sequence features found in collagen and/or elastin comprise an octapeptide.
  • the peptide is an elastin-derived peptide.
  • the elastin-derived peptide comprises an amino acid sequence XGVXXG.
  • the elastin-derived peptide comprises a sequence according to amino acid sequence IGVAPG, VGVAPG, or VGVTAG.
  • compositions and methods described herein further comprise one or more tripeptides.
  • compositions described herein comprise an octapeptide.
  • the octapeptide is provided at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %), relative to the total weight of the composition.
  • the octapeptide is provided in a range of about 0.00001 wt.% - 1.0 wt.%, 0.0001 wt.% - 0.01 wt.%, about 0.0001 wt.% to 0.001 wt.%, about 0.0001 wt.% to 0.0005 wt.%, about 0.25% to about 10%, about 0.5% to about 8%, about 0.75% to about 6%, or about 1% to about 4% by weight.
  • the octapeptide is provided in a concentration of at least about 0.10, 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 parts per million (ppm).
  • the octapeptide is provided in a concentration of about 0.1 ppm to about 200 ppm, about 1 ppm to about 200 ppm, about 1 to about 150 ppm, about 1 to about 100 ppm, about 1 to about 100 ppm, about 1 to about 50 ppm, about 1 to about 25 ppm, about 1 to about 20 ppm, about 1 to about 10 ppm, or about 10 to about 100 ppm. In some embodiments, the octapeptide is provided in a concentration of about 10 ppm.
  • the octapeptide may be present in an amount of from about 50 ppm or less to about 100, 200, 300, 400, or 500 ppm or more, e.g., 50 ppm to 150 ppm.
  • the hexapeptide is typically present in an amount of from about 50 ppm or less to about 100, 200, 300, 400, or 500 ppm or more, e.g., 50 ppm to 150 ppm.
  • the octapeptide is provided in a concentration of at least about 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 pg /mL.
  • the octapeptide is provided in a concentration of about 1 to about 200 pg/mL, about 1 to about 150 pg/mL, about 1 to about 100 pg/mL, about 1 to about 50 pg/mL, about 1 to about 10 pg /mL, about 10 to about 100 pg /mL, about 10 to about 50 pg/mL, about 20 to about 100 pg/mL, or about 30 to about 100 pg /mL. In some embodiments, the octapeptide is provided in a concentration of about 10 pg/mL.
  • the octapeptide is provided in a concentration of about 10 pg/mL to 20 pg/mL. In some embodiments, the octapeptide is provided in a concentration of about 20 pg/mL. In some embodiments, the octapeptide is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 microgram per milliliter.
  • compositions as described herein comprise a varying concentration of peptide.
  • a peptide is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 ppm.
  • a peptide is present in a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 ppm.
  • a peptide is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or more than 1000 microgram per milliliter (pg/mL). In some instances, a peptide is present in a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 microgram per milliliter.
  • a peptide is present from about 0.01% to about 10%, about 0.01% to about 0.02%, about 0.01% to about 0.03%, about 0.01% to about 0.04%, about 0.01% to about 0.05%, about 0.01% to about 0.1%, about 1% to about 5%, or about 1% to about 10% by weight (wt. %).
  • compositions as described herein comprise one or more peptides.
  • a peptide of the one or more peptides is present at about 50 ppm or less to 1000, 5000, 10000, 50000, 100000, 500000 ppm or more, e.g., 100 ppm of the peptide, or any other suitable amount.
  • a peptide of the one or more peptides is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or more than 1000 ppm.
  • a peptide of the one or more peptides is present in a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 ppm. In some instances, a peptide of the one or more peptides is present at about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or more than 1000 microgram per milliliter (pg /mL).
  • a peptide of the one or more peptides is present in a range of about 1 to about 100, about 1 to about 50, about 1 to about 40, about 1 to about 30, about 1 to about 20, about 1 to about 10, about 5 to about 90, about 10 to about 80, about 20 to about 60, or about 30 to about 50 microgram per milliliter. In some instances, a peptide of the one or more peptides is present from about 0.01% to about 10%, about 0.01% to about 0.02%, about 0.01% to about 0.03%, about 0.01% to about 0.04%, about 0.01% to about 0.05%, about 0.01% to about 0.1%, about 1% to about 5%, or about 1% to about 10% by weight (wt. %).
  • a peptide of the one or more peptides is provided at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %).
  • a peptide of the one or more peptides is provided in a range of about 0.25% to about 10%, about 0.5% to about 8%, about 0.75% to about 6%, or about 1% to about 4% by weight. In some embodiments, each peptide of the one or more peptides is provided in a range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 3%, or about 0.02% to about 2% by weight.
  • the tripeptide is a synthetic tripeptide.
  • the synthetic tripeptide is tetradecyl aminobutyroylvalylaminobutyric urea trifluoroacetate.
  • the tripeptide is typically present in an amount of from about 50 ppm or less to about 100, 200, 300, 400, or 500 ppm or more, e.g., 50 ppm to 150 ppm.
  • the hexapeptide is typically present in an amount of from about 50 ppm or less to about 100, 200, 300, 400, or 500 ppm or more, e.g., 50 ppm to 150 ppm.
  • the tripeptide is tripeptide-1.
  • the tripeptide-1 is provided at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %).
  • the tripeptide-1 is provided in a range of about about 0.01 wt.% to about 2 wt.%, about 0.25% to about 10%, about 0.5% to about 8%, about 0.75% to about 6%, or about 1% to about 4% by weight. In some embodiments, the tripeptide-1 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 ppm. In some embodiments, the tripeptide-1 is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 ppm.
  • the tripeptide-1 is provided in a range of about 1 to about 10 ppm. In some embodiments, the tripeptide-1 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 microgram per milliliter (pg/mL). In some embodiments, the tripeptide-1 is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 microgram per milliliter.
  • the elastin-derived peptide is a synthetic peptide.
  • the elastin-derived peptide is a peptide that comprises elastogenic potential that mimics the elastin-derived VGVAPG, IGVAPG, VGVTAG (identified in the IGF-l-binding protein- 1 (IGFBP-1)), or derivatives thereof.
  • the elastin-derived peptide is a synthetic peptide that comprises elastogenic potential that mimics the elastin-derived VGVAPG, IGVAPG, VGVTAG (identified in the IGF-l-binding protein-1 (IGFBP-1)), or derivatives thereof.
  • the elastin-derived peptide comprises an amino acid sequence XGVXXG. In some embodiments, the elastin-derived peptide comprises a sequence according to amino acid sequence IGVAPG, VGVAPG, or VGVTAG. In some embodiments, the synthetic peptide is a hexapeptide. In some embodiments, the hexapeptide is hexapeptide-11. In some embodiments, hexapeptide-11 is encapsulated in a liposome. In some embodiments, tripeptide is encapsulated in a liposome. In some embodiments, octapeptide is encapsulated in a liposome.
  • octapeptide-45 is encapsulated in a liposome.
  • TriHex and octapeptide-45 are encapsulated in a liposome.
  • the synthetic peptide is a lactoferrin.
  • lactoferrin is encapsulated in a liposome.
  • encapsulation in a liposome increases the efficiency of peptide delivery to cells.
  • encapsulation in a liposome increases the efficiency of peptide delivery to cells in deeper layers of the skin.
  • encapsulation in a liposome increases the efficiency of peptide delivery to cells in deeper epidermal layers of the skin.
  • encapsulation in a liposome increases the efficiency of peptide delivery to cells in deeper dermal layers of the skin. In some embodiments, encapsulation in a liposome increases half-life of a peptide in a formulation. In some embodiments, encapsulation in a liposome increases half-life of a tripeptide in a formulation. In some embodiments, encapsulation in a liposome increases half-life of a hexapeptide-11 in a formulation. In some embodiments, encapsulation in a liposome increases half-life of an octapeptide in a formulation.
  • encapsulation in a liposome increases half-life of octapeptide-45 in a formulation. In some embodiments, encapsulation in a liposome yields a formulation having a sustained release of a peptide upon application. In some embodiments, encapsulation in a liposome yields a formulation having a sustained release of a peptide upon topical application. In some embodiments, encapsulation in a liposome yields a formulation having a sustained release of a peptide upon topical application to facial skin.
  • the elastin-derived peptide is provided at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %).
  • the elastin-derived peptide is provided in a range of about 0.00001% to about 10%, about 0.0001 % to about 0.01 %, about 0.0003% to about 9%, about 0.0005% to about 8%, about 0.001% to about 4%, or about 0.001% to 2% by weight (wt. %), or any range or value therein between.
  • the elastin-derived peptide is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 ppm.
  • the elastin-derived peptide is provided in a range of about 1 to about 10 ppm.
  • the elastin-derived peptide is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 ppm. In some embodiments, the elastin-derived peptide is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 microgram per milliliter (pg/mL). In some embodiments, the elastin-derived peptide is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 microgram per milliliter.
  • the hexapeptide- 12 is provided at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %).
  • the hexapeptide- 12 is provided in a range of about 0.00001% to about 10%, about 0.0001% to about 0.01%, about 0.0003%, to about 9%, about 0.0005% to about 8%, or about 0.001% to about 4%, about 0.001% to about 2% by weight (wt. %), or any range or value therein between.
  • the hexapeptide- 12 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 ppm.
  • the hexapeptide- 12 is provided in a range of about 1 to about 10 ppm.
  • the hexapeptide- 12 is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 ppm. In some embodiments, the hexapeptide- 12 is provided at least or about 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or more than 25 microgram per milliliter (pg /mL). In some embodiments, the hexapeptide- 12 is provided in a range of about 0.25 to about 10, about 0.5 to about 8, about 1 to about 6, or about 2 to about 4 microgram per milliliter.
  • compositions described herein comprise hexapeptide-11.
  • the hexapeptide-11 is provided at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %).
  • the hexapeptide-11 is provided in a range of about 0.00001% to about 10%, about 0.0001% to about 0.01%, about 0.0003% to about 8%, about 0.0005%, to about 6%, about 0.001% to about 4%, about 0.001% to about 2%, about 0.005% to about 2%, or about 0.01% to about 1% by weight (wt. %), or any range or value therein between.
  • the hexapeptide-11 is provided in a range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 3%, or about 0.02% to about 2%.
  • the hexapeptide 11 is provided in a range of about 0.005% to about 0.02% by weight. In some embodiments, the hexapeptide-11 is provided at least or about 0.1 ppm, 3 ppm, 5 ppm, 10 ppm, 50 ppm, 55 ppm, 500 ppm, 1,000 ppm, 2,500 ppm, 5,000 ppm, or more than 5,000 ppm. In some embodiments, the hexapeptide-11 is provided in a range of about 5 ppm to about 100 ppm, about 10 ppm to about 1000 ppm, about 50 ppm to about 1500 ppm, or about 500 ppm to about 5,000 ppm.
  • the hexapeptide-11 is about 1000 ppm. In some embodiments, the hexapeptide-11 is provided at least or about 5, 10, 20, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, or more than 500 microgram per milliliter (pg /mL). In some embodiments, the hexapeptide-11 is provided in a range of about 25 to about 250, about 50 to about 200, about 75 to about 150, about 200 to about 300, or about 200 to about 400 microgram per milliliter.
  • the peptide can be functionalized.
  • the peptide can be functionalized with a fatty acid, e.g., myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, a-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, caprylic acid, capric acid, lauric acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, or the like.
  • a fatty acid e.g., myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid,
  • Palmitoyl or myristoyl functionalization can be desirable in certain embodiments as it exhibits enhanced penetration when compared to other fatty acids.
  • the peptide is functionalized with a chemical group.
  • the peptide is functionalized with acetyl.
  • the peptide is functionalized with a functional group comprising no more than 14 carbons.
  • the peptide is functionalized with a functional group comprising no more than 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more than 20 carbons. In some instances, the peptide is non-palmitoylated. Without wishing to be limited to a particular theory, incorporation of the peptide in a liposome, in some embodiments, increases the lipophilicity of a peptide that is functionalized or is not functionalized.
  • GHK glycine-histidine-lysine
  • GHK is a peptide sequence that is rarely found in the class of proteins in general, but is frequently found in extracellular matrix proteins. The small size of GHK permits it to approach membrane receptors far more easily than larger peptides. Further, its unique, copper-binding structure enhances copper transport into and out of cells and promotes wound healing through several different but related pathways. Due to its strong copper binding structure, GHK can be provided in the form of GHK-Cu (copper-bound GHK form).
  • compositions described herein comprise an octapeptide.
  • the octapeptide comprises an amino acid sequence GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8).
  • the octapeptide comprises an amino acid sequence GDGDGASA (SEQ ID NO: 1).
  • the octapeptide comprises an amino acid sequence GPMGPSGP (SEQ ID NO: 2). In some embodiments, the octapeptide comprises an amino acid sequence GLGPGARA (SEQ ID NO: 3). In some embodiments, the octapeptide comprises an amino acid sequence GPQGFQGP (SEQ ID NO: 4). In some embodiments, the octapeptide comprises an amino acid sequence GPHGVREA (SEQ ID NO: 5). In some embodiments, the octapeptide comprises an amino acid sequence GPMGPRGP (SEQ ID NO: 6). In some embodiments, the octapeptide comprises an amino acid sequence GPGKNGDD (SEQ ID NO: 7). In some embodiments, the octapeptide comprises an amino acid sequence GPMGPRGP (SEQ ID NO: 8). In some embodiments, octapeptide-45 is GPHGVREA (SEQ ID NO: 5).
  • the octapeptide is a synthetic peptide. In some embodiments, the octapeptide is a peptide that stimulates HA synthesis. In some embodiments, the octapeptide is a peptide that stimulates high molecular weight hyaluronic acid (high MW HA) synthesis. In some embodiments, the octapeptide is a peptide that stimulates high MW HA synthesis in the dermis. In some embodiments, the octapeptide is a peptide that stimulates high MW HA synthesis in the skin.
  • high MW HA high molecular weight hyaluronic acid
  • the octapeptide comprises an octapeptide having an amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD.
  • the octapeptide comprises GPHGVREA.
  • the octapeptide is GPHGVREA.
  • the octapeptide comprises a chemical structure of octapeptide-45 (FIG. 22).
  • compositions according to the present disclosure comprise liposomes for improved distribution, efficacy, bioavailability, and/or activity.
  • Liposomal compositions may improve distribution, efficacy, bioavailability, and/or activity of the active ingredient by improving delivery and tissue (e.g., skin) penetration.
  • improved delivery and skin penetration result from the active ingredient being incorporated e.g., encapsulated) in a liposome.
  • the active ingredient is one or more peptides encapsulated in a liposome.
  • Liposomal compositions as described herein may comprise a peptide encapsulated in a liposome.
  • the peptide encapsulated in the liposome is selected from the group consisting of hexapeptide- 12, tripeptide-1, one or more octapeptides (e.g., octapeptide-45), hexapeptide- 11, hexapeptide-38, tetrapeptide-2, or any combination thereof.
  • the peptide or peptides encapsulated in a liposome are functionalized with an acetyl group, a palmitoyl group, a myristoyl group, or any combination thereof.
  • Liposomal compositions as described herein may comprise various ingredients encapsulated in a liposome.
  • the ingredient is lactoferrin.
  • the ingredient is phosphatidylserine.
  • the ingredient is Ledum palustre extract.
  • the ingredient is Arnica montana extract.
  • the ingredient is sodium hyaluronate.
  • the ingredient is larger than 50 kDa.
  • Lecithin and other phospholipids may be used to prepare liposomes containing the peptide compositions as described herein.
  • liposomes are used to prepare one or more peptides.
  • the peptide is functionalized with an acetyl group. Formation of lipid vesicles occurs when phospholipids such as lecithin are placed in water and consequently form one bilayer or a series of bilayers, each separated by water molecules, once enough energy is supplied. Liposomes can be created by sonicating phospholipids in water. Low shear rates create multilamellar liposomes. Continued high-shear sonication tends to form smaller unilamellar liposomes. Hydrophobic chemicals can be dissolved into the phospholipid bilayer membrane. The lipid bilayers of the liposomes deliver the peptide compositions as described herein.
  • the phospholipids used to prepare the liposomal compositions described herein may comprise a transition phase temperature of about 10 °C to about 25 °C.
  • the phospholipids comprise a transition phase temperature of about 10 °C, 12 °C, 14 °C, 16 °C, 18 °C, 20 °C, 22 °C, 24 °C, 26 °C, 28 °C, 30 °C, 32 °C, 34 °C, 36 °C, 38 °C, 40 °C, or more than 40 °C.
  • the phospholipids comprise a transition phase temperature in a range of about 10 °C to about 40 °C, about 12 °C to about 36 °C, about 14 °C to about 32 °C, about 16 °C to about 20 °C, or about 21 °C to about 25 °C.
  • the topical composition may contain micelles, or an aggregate of surfactant molecules dispersed in an aqueous solution.
  • Micelles may be prepared by dispersing an oil solvent in an aqueous solution comprising a surfactant, where the surfactant concentration exceeds the critical micelle concentration.
  • the resulting composition contains micelles, i.e., spherical oil droplets.
  • the liposomal composition may contain micelles, or an aggregate of surfactant molecules dispersed in an aqueous solution.
  • Micelles may be prepared by dispersing an oil solvent in an aqueous solution comprising a surfactant, where the surfactant concentration exceeds the critical micelle concentration.
  • the resulting formulation contains micelles, i.e., spherical oil droplets surrounded by a membrane of polar surfactant molecules, dispersed in the aqueous solvent.
  • the contacting occurs at a temperature of about 10 °C, 12 °C, 14 °C, 16 °C, 18 °C, 20 °C, 22 °C, 24 °C, 26 °C, 28 °C, 30 °C, 32 °C, 34 °C, 36 °C, 38 °C, 40 °C, or more than 40 °C. In some instances, the contacting occurs at a temperature in a range of about 10 °C to about 40 °C, about 12 °C to about 36 °C, about 14 °C to about 32 °C, about 16 °C to about 20 °C, or about 21 °C to about 25 °C.
  • Methods for preparing a composition comprising a peptide encapsulated in a liposome may comprise use of a solvent.
  • the solvent is water.
  • the solvent is an organic solvent.
  • Exemplary organic solvents include, but are not limited to, petroleum ether, cyclohexane, toluene, carbon tetrachloride, dichloromethane, chloroform, diethyl ether, diisopropyl ether, ethyl acetate, butanol, n-propanol, ethanol, methanol, polyethylene glycol, propylene glycol, and pyridine.
  • the solvent is a glycol.
  • the solvent is butylene glycol.
  • the solvent is caprylyl glycol.
  • the solvent is propanediol (propylene glycol).
  • the solvent may be used at various percentages. In some instances, the solvent is provided at least or about 0.001%, 0.005%, 0.01%, 0.02%, 0.05%, 0.10%, 0.20%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%
  • the solvent may be propanediol, butylene glycol, or caprylyl glycol.
  • Methods as described herein comprises combining the peptide and a solvent to form a mixture; and contacting the mixture with an aqueous solution comprising liposomes, wherein the aqueous solution comprises a percentage of water and a percentage of liposomes.
  • the aqueous solution comprises at least or about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more than 90% water.
  • the aqueous solution comprises water in a range of about 10% to about 95%, about 20% to about 90%, about 30% to about 85%, about 40% to about 80%, or about 50% to about 60%.
  • the aqueous solution comprises at least or about 20%, 30%, 40%, 50%, 60%, or more than 60% liposomes. In some instances, the aqueous solution comprises liposomes in a range of about 10% to about 80%, about 20% to about 70%, or about 30% to about 60%.
  • a ratio of liposomes to water may be in a range of about 1 :9 to about 3:7. In some instances, the ratio of liposomes to water may be at least or about 1 : 10, 1 :9, 1 :8, 1 :7, 1 :6, 1 :5, 1 :4, 1 :3, or 1 :2.
  • Methods for generation of liposomal compositions as described herein may result in an entrapment efficacy of no more than 100%.
  • the entrapment efficacy is no more than 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 99.5%.
  • the peptide or peptides are provided at least or about 0.00001%, 0.00003%, 0.00005%, 0.0001%, 0.0003%, 0.0005%, 0.001%, 0.003%, 0.005%, 0.0055%, 0.01%, 0.03%, 0.05%, 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% of the composition.
  • the peptide or peptides are provided at least or about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 22%, 24%, 26%, 28%, 30% or more than 30% of the composition. In some embodiments, the peptide or peptides are provided in a range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 5%, or about 0.02% to about 2% by weight. In some embodiments, the peptide is provided at about 0.03% of the composition.
  • liposomal compositions wherein the liposomes comprise a percentage of the composition.
  • the liposomes are provided at least or about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 22%, 24%, 26%, 28%, 30% or more than 30% of the composition.
  • the liposomes are provided in a range of about 5% to about 90%, about 10% to about 80%, about 20% to about 70%, about 30% to about 60%, about 10% to about 30%, or about 20% to about 40%.
  • the liposomes are provided at about 30%.
  • the liposomes are provided at 27%.
  • liposomal compositions as described herein comprise an average particle size of at most 220 nanometers (nm).
  • the average particle size is at most 100 nm, 105 nm, HO nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, 230 nm, 240 nm, 250 nm, 260 nm, 270 nm, 280 nm, 290 nm, 300 nm, 320 nm, 340 nm, 360 nm, 380
  • the average particle size is about 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, 230 nm, 240 nm, 250 nm, 260 nm, 270 nm, 280 nm, 290 nm, 300 nm, 320 nm, 340 nm, 360 nm, 380 nm, or 400 nm.
  • the average particle size is in a range of about 50 nm to about 500 nm, about 100 nm to about 400 nm, about 150 nm to about 220 nm, about 180 nm to about 220 nm, or about 190 nm to about 210 nm.
  • the liposomal compositions comprise an active agent that has a molecular weight of no more than about 600 Daltons (Da).
  • the active agent has a molecular weight of at least or about 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975, 1000, or more than 1000 Daltons (Da).
  • the active agent has a molecular weight of at least or about 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 4000, 5000, 6000, or more than 6000 Daltons (Da).
  • the active agent has a molecular weight in a range of about 50 to about 1000, about 100 to about 900, about 200 to about 800, about 300 to about 700, or about 400 to about 600 Daltons (Da).
  • the active agent is a peptide or peptides as disclosed herein.
  • a poly dispersity index (Pdl) of a liposomal composition as described herein in some embodiments, is in a range of 0 to about 0.2. In some instances, the polydispersity index is about 0.01, 0.025, 0.05, 0.1, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, or 0.8. In some instances, the poly dispersity index is in a range of about 0.01 to about 0.8, about 0.025 to about 0.75, about 0.05 to about 0.6, or about 0.1 to about 0.3.
  • an intercept of a liposomal composition as described herein is in a range of about 0.85 to about 0.95. In some instances, the intercept is the amplitude. In some instances, the intercept is at least or about 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, or 0.95.
  • the liposomes comprise propanediol, lecithin, or a combination thereof.
  • the propanediol is provided at least or about 0.001%, 0.005%, 0.01%, 0.02%, 0.05%, 0.10%, 0.20%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %).
  • the propanediol is provided in a range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 3%, or about 0.02% to about 2% by weight.
  • the lecithin is provided at least or about 0.001%, 0.005%, 0.01%, 0.02%, 0.05%, 0.10%, 0.20%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %), relative to the total weight of the composition.
  • the lecithin is provided in a range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 3%, or about 0.02% to about 2% by weight, relative to the total weight of the composition.
  • the liposomes comprise propanediol and lecithin.
  • the propanediol and lecithin are provided at least or about 0.001%, 0.005%, 0.01%, 0.02%, 0.05%, 0.10%, 0.20%, 0.25%, 0.50%, 0.75%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 8%, 9%, 10%, or more than 10% by weight (wt. %), relative to the total weight of the composition.
  • the propanediol and lecithin are provided in a range of about 0.001% to about 6%, about 0.002% to about 4%, about 0.01% to about 3%, or about 0.02% to about 2% by weight, relative to the total weight of the composition. In some embodiments, the propanediol and lecithin are provided at about 0.90% by weight, relative to the total weight of the composition.
  • Described herein are liposomal compositions comprising improved distribution, efficacy, bioavailability, and/or activity.
  • the liposomal compositions may comprise improved distribution, efficacy, bioavailability, and/or activity as compared to compositions not comprising liposomes.
  • the distribution is improved by at least or about 0.5X, LOX, 1.5X, 2. OX, 2.5X, 3. OX, 4. OX, 4.5X, 5X, or more than 5X as compared to compositions not comprising liposomes.
  • the efficacy is improved by at least or about 0.5X, LOX, 1.5X, 2. OX, 2.5X, 3. OX, 4. OX, 4.5X, 5X, or more than 5X as compared to compositions not comprising liposomes.
  • the bioavailability is improved by at least or about 0.5X, LOX, 1.5X, 2. OX, 2.5X, 3. OX, 4.
  • the activity is improved by at least or about 0.5X, LOX, 1.5X, 2. OX, 2.5X, 3. OX, 4. OX, 4.5X, 5X, or more than 5X as compared to compositions not comprising liposomes.
  • the distribution, efficacy, bioavailability, and/or activity may be improved by at least or about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than 90% as compared to compositions not comprising liposomes.
  • Liposomal compositions and methods as described herein, in some embodiments, are topical compositions.
  • the liposomal compositions are oil free.
  • the liposomal compositions are preservative free.
  • the liposomal formulation is an aqueous formulation.
  • the liposomal formulation is an anhydrous formulation.
  • the liposomal composition comprises a pH in a range of about 5 to about 8. In some instances, the liposomal composition comprises a pH of at least or about 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • Methods and compositions as described herein may result in improved follicular penetration.
  • the follicular penetration is improved by at least or about 0.5X, LOX, 1.5X, 2. OX, 2.5X, 3. OX, 4. OX, 4.5X, 5X, or more than 5X.
  • the follicular penetration may be improved by at least or about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or more than 90%.
  • compositions result in follicular penetration of a depth of at least or about 0.5, 0.75, 1, 1.25, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 7, 8, 9, 10, or more than 10 millimeters.
  • compositions described herein alter gene expression in skin cells.
  • exposure of dermal cells e.g., dermal fibroblasts
  • an octapeptide results in upregulation of HAS2 expression within dermal cells.
  • the dermal cells comprise dermal fibroblasts.
  • the octapeptide comprises one or more octapeptides with the amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD.
  • the octapeptide is octapeptide-45.
  • hexapeptide e.g., hexapeptide- 11
  • exposure of dermal cells (e.g., dermal fibroblasts) or epidermal cells (e.g., keratinocytes) to a hexapeptide results in upregulation of HAS2 expression.
  • exposure of dermal cells (e.g., dermal fibroblasts) or epidermal cells (e.g., keratinocytes) to a tripeptide and a hexapeptide (TriHex - hexapeptide- 12 + tripeptide- 1) results in upregulation of HAS2 expression.
  • epidermal cells e.g., keratinocytes
  • a hexapeptide e.g., hexapeptide-11
  • exposure of epidermal keratinocytes to TriHex results in upregulation of HAS2 expression.
  • exposure of epidermal keratinocytes to hexapeptide-11 results in upregulation of HAS2 expression.
  • epidermal cells e.g., keratinocytes
  • a hexapeptide e.g., hexapeptide-11
  • exposure of epidermal keratinocytes to hexapeptide-11 results in downregulation of HYAL2 expression.
  • exposure of epidermal cells to a hexapeptide or octapeptide results in upregulation of EGR3 expression.
  • exposure of epidermal (e.g., keratinocytes) cells to tripeptide-1 and hexapeptide- 12 (TriHex) results in upregulation of EGR3 expression.
  • exposure of epidermal cells to hexapeptide-11 results in upregulation of EGR3 expression.
  • exposure of epidermal keratinocytes to hexapeptide results in upregulation of EGR3 expression.
  • exposure of epidermal keratinocytes to hexapeptide-11 results in upregulation of EGR3 expression.
  • exposure of epidermal keratinocytes to an octapeptide results in upregulation of EGR3 expression.
  • exposure of human dermal fibroblasts to TriHex results in upregulation of LAMA3 expression.
  • exposure of human dermal fibroblasts to TriHex results in upregulation of MERTK expression.
  • exposure of human dermal fibroblasts to an octapeptide results in upregulation of SI 00 A2 expression.
  • exposure of human dermal fibroblasts to an octapeptide results in upregulation of LAMA3 expression.
  • exposure of human dermal fibroblasts to an octapeptide (e.g., octapeptide- 45) results in upregulation of MERTK expression.
  • exposure of human dermal fibroblasts to an octapeptide results in upregulation of ITGB4 expression.
  • exposure of human dermal fibroblasts to TriHex + octapeptide results in upregulation of SI 00 A2 expression.
  • exposure of human dermal fibroblasts to TriHex + octapeptide results in synergistic upregulation of S100A2 expression, when compared to expression after exposure to TriHex alone or octapeptide alone.
  • exposure of human dermal fibroblasts to TriHex + octapeptide results in upregulation of LAMA3 expression.
  • exposure of human dermal fibroblasts to TriHex + octapeptide results in synergistic upregulation of LAMA3 expression, when compared to expression after exposure to TriHex alone or octapeptide alone.
  • exposure of human dermal fibroblasts to TriHex + octapeptide results in upregulation of MERTK expression.
  • exposure of human dermal fibroblasts to TriHex + octapeptide results in synergistic upregulation of MERTK expression, when compared to expression after exposure to TriHex alone or octapeptide alone.
  • exposure of human dermal fibroblasts to TriHex + octapeptide results in upregulation of ITGB4 expression.
  • octapeptide e.g., octapeptide-45
  • exposure of human dermal fibroblasts to TriHex + octapeptide results in synergistic upregulation of ITGB4 expression, when compared to expression after exposure to TriHex alone or octapeptide alone.
  • the composition comprising an octapeptide results in increase synthesis of high MW HA as compared to a skin untreated with the composition.
  • the octapeptide in the composition stimulates synthesis of high MW HA.
  • the increase in HA synthesis is by stimulation of hyaluronic acid synthase (HAS) activity.
  • HAS comprises HAS2.
  • the increase in HA synthesis is by downregulation of hyaluronidase expression.
  • the hyaluronidase comprises hyaluronidase 2 (HYAL2).
  • the octapeptide stimulates HAS2 in fibroblasts, enzyme that stimulates HA production. In some embodiments, the octapeptide is encapsulated into a phospholipid delivery system for increased absorption and activity. In some embodiments, the hexapeptide provides upregulation of HAS2 in keratinocytes and potent downregulation of HYAL2 in keratinocytes. In some embodiments, the compositions described herein stimulate the synthesis of HA in dermal and/or epidermal cells. In some embodiments, the HA synthesized with the application of the composition comprises high MW HA than the HA synthesized without the composition.
  • the high MW HA produced by stimulation has a MW of at least 0.5 MDa, 1.0 MDa, 1.5 MDa, or 2.0 MDa. In some embodiments, the high MW HA produced by stimulation has a MW of about 1 MDa to 4 MDa.
  • exposure of dermal cells to octapeptide stimulates the production of HA. In some embodiments, exposure of dermal cells to octapeptide-45 stimulates the production of HA. In some embodiments, exposure of dermal fibroblasts to octapeptide stimulates the production of HA.
  • exposure of dermal fibroblasts to octapeptide at a concentration of at least about 100 pg/ml stimulates the production of HA. In some embodiments, exposure of epidermal cells to octapeptide stimulates the production of HA. In some embodiments, exposure of epidermal cells to octapeptide-45 stimulates the production of HA. In some embodiments, exposure of epidermal keratinocytes to octapeptide stimulates the production of HA. In some embodiments, exposure of epidermal keratinocytes to octapeptide-45 stimulates the production of HA.
  • exposure of dermal fibroblasts to lactoferrin at a concentration of at least about 500 pg/ml stimulates the production of HA.
  • exposure of dermal fibroblasts to phosphatidylserine at a concentration of at least about 500 pg/ml stimulates the production of HA.
  • exposure of dermal fibroblasts to Tremella at a concentration of at least about 500 pg/ml stimulates the production of HA.
  • exposure of dermal fibroblasts to hydroxymethoxphenyl decanone at a concentration of at least about 250 pg/ml stimulates the production of HA.
  • exposure of skin to the compositions comprising one or more octapeptides increases expression of CD44.
  • exposure of skin to the composition comprising octapeptide-45 increases expression of CD44.
  • exposure of epidermal keratinocytes and/or dermal fibroblasts to the composition increases the expression of CD44 in the exposed cells.
  • increases in expression of CD44 contribute to the remodeling of dermal ECM.
  • increases in expression of CD44 contribute to the remodeling of dermal ECM to reverse the appearance of solar elastosis.
  • increases in expression of CD44 contribute to a reduction in skin redness.
  • increases in expression of CD44 contribute to a reduction in skin inflammation. In some embodiments, increases in expression of CD44 contribute to a reduction in the appearance of wrinkles. In some embodiments, increases in expression of CD44 contribute to a reduction in the appearance of fine lines. In some embodiments, increases in expression of CD44 contribute to a reduction in the appearance of deep lines. In some embodiments, increases in expression of CD44 contribute to a reduction in the appearance of deep lines in facial skin. In some embodiments, increases in expression of CD44 contribute to a reduction in the appearance of crepiness. In some embodiments, increases in expression of CD44 contribute to a reduction in the appearance of pore size. In some embodiments, increases in expression of CD44 contribute to an improvement in the appearance of skin texture. In some embodiments, increases in expression of CD44 contribute to a reduction in skin dryness. In some embodiments, increases in expression of CD44 contribute to an improvement in skin hydration.
  • a formulation for use on the skin comprises a hyaluronic acid or a derivative thereof.
  • the hyaluronic acid comprises high molecular weight hyaluronic acid (high MW HA), or a derivative thereof.
  • the high MW HA comprises high MW HA having an average molecular weight of at least about 0.5 MDa, 1.0 MDa, 1.5 MDa, 2.0 MDa, 3.0 MDa, 4.0 MDa, or 5.0 MDa.
  • the high MW HA comprises high MW HA of a molecular weight of about 1 MDa to about 5 MDa.
  • high MW HA comprises a sodium hyaluronate.
  • high MW HA comprises a sodium hyaluronate crosspolymer.
  • compositions comprising phosphatidylserine.
  • compositions comprising Tremella fuciformis extract are compositions comprising hydroxymethoxyphenyl decanone.
  • a composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyl decanone, or combinations thereof.
  • the composition provided herein increases production of high MW HA in skin cells.
  • the composition increases production of high MW HA in epidermal cells. In some embodiments, the composition increases production of high MW HA in dermal cells. In some embodiments, the composition provided herein increases the integrity of the barrier beneath the stratum basale. In some embodiments, the composition increases the thickness of the barrier beneath the stratum basale. In some embodiments, the composition increases expression of genes regulating the barrier function of the stratum basale. In some embodiments, the composition increases the barrier function of skin. In some embodiments, the composition increases the barrier function of skin to prevent excessive water loss. In some embodiments, the composition increases the barrier function of skin to maintain a higher moisture content. In some embodiments, the composition increases the barrier function of skin to increase skin hydration.
  • the composition increases the barrier function of skin to reduce skin dryness. In some embodiments, the composition increases the barrier function of the skin to reduce redness. In some embodiments, the composition increases the barrier function of the skin to reduce inflammation. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of wrinkles. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of fine lines. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of deep lines in skin. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of deep lines in facial skin. In some embodiments, the composition increases the barrier function of the skin to reduce the appearance of crepiness.
  • the composition increases the barrier function of the skin to reduce the appearance of pore size. In some embodiments, the composition increases the barrier function of the skin to improve the appearance of skin texture. In some embodiments, the composition is aqueous. In some embodiments, the composition reduces inflammation as assessed by a reduction or a lack of increase in an expression level of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL- 12b), or cluster of differentiation-80 (CD80), or combinations thereof. In some embodiments, the composition increases CD44 expression. In some embodiments, the composition reduces redness of a skin of an individual after applying the composition.
  • NOS2 nitric oxide synthase 2
  • TNF tumor necrosis factor
  • IL- 12b interleukin 12
  • CD80 cluster of differentiation-80
  • the composition reduces redness of a skin of an individual after applying the composition and the reduction in redness is measured by photography.
  • the composition increases skin hydration in a skin of an individual after applying the composition.
  • the composition increases skin hydration in a skin of an individual after applying the composition.
  • the composition promotes HA synthesis.
  • the composition upregulates hyaluronic acid synthase (HAS) expression.
  • the composition upregulates hyaluronic acid synthase (HAS) expression, wherein the HAS comprises HAS2.
  • the composition downregulates hyaluronidase expression.
  • the composition downregulates hyaluronidase expression wherein the hyaluronidase comprises hyaluronidase 2 (HYAL2).
  • the composition comprises tripeptide-1, hexapeptide- 12, and octapeptide-45.
  • the composition increases elastin production.
  • the composition increases production of tropoelastin.
  • the composition increases production of microfibrillar-associated protein 4 (MF AIM).
  • MF AIM microfibrillar-associated protein 4
  • the composition upregulates SAI 002 expression.
  • the composition upregulates LAMA3 expression.
  • the composition upregulates MERTK expression.
  • the composition upregulates ITGB4 expression.
  • the composition upregulates gene expression for one or more pathways related to ECM regulation.
  • the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyl decanone, or combinations thereof.
  • the composition provided herein increases production of high MW HA in skin cells.
  • the composition increases production of high MW HA in epidermal cells.
  • the composition increases production of high MW HA in dermal cells.
  • the compositions provided herein decrease one or more of the appearance of redness, skin inflammation, the appearance of rosacea, the appearance of erythema, the appearance of fine blood vessels in skin, the appearance of wrinkles, the appearance of fine lines, the appearance of deep lines, the appearance of crepiness of skin, the appearance of pore size, excess water loss in the skin, dryness of the skin, or the appearance of solar estastosis.
  • the compositions provided herein improve the appearance of skin texture and/or increase skin hydration.
  • the compositions provided herein increase the rate of collagen turnover.
  • the collagen turnover comprises replacing old thin collagen fibers in dermal ECM into denser, healthier collagen fibers in dermal ECM.
  • compositions for reducing inflammation comprising an octapeptide; and a hyaluronic acid (HA) or derivatives thereof, wherein the composition reduces inflammation.
  • HA or derivatives thereof has a molecular weight of at least 0.5 MDa.
  • the HA or derivatives thereof has a molecular weight of at least 1 MDa.
  • the high MW HA or derivatives comprises sodium hyaluronate.
  • the sodium hyaluronate comprises a sodium hyaluronate crosspolymer.
  • the octapeptide comprises at least one of an amino acid sequence GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8).
  • the octapeptide comprises an amino acid sequence GPHGVREA (SEQ ID NO: 5).
  • the octapeptide is the amino acid sequence GPHGVREA (SEQ ID NO: 5).
  • the octapeptide comprises octapeptide-45.
  • the composition comprises a synthetic tripeptide.
  • the synthetic tripeptide is tetradecyl aminobutyroylvalylaminobutyric urea trifluoroacetate.
  • the composition comprises a hexapeptide.
  • the hexapeptide is hexapeptide-11.
  • the hexapeptide-11 is encapsulated in a liposome.
  • the composition comprises lactoferrin. In some embodiments, the lactoferrin is encapsulated in a liposome.
  • the composition comprises phosphatidylserine. In some embodiments, the composition comprises Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyl decanone. In some embodiments, the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyl decanone, or combinations thereof. In some embodiments, the composition is aqueous.
  • the reduction in inflammation is assessed by a reduction or a lack of increase in an expression level of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL- 12b), or cluster of differentiation-80 (CD80), or combinations thereof.
  • NOS2 nitric oxide synthase 2
  • TNF tumor necrosis factor
  • IL- 12b interleukin 12
  • CD80 cluster of differentiation-80
  • the composition increases CD44 expression.
  • the composition reduces redness of a skin of an individual after applying the composition.
  • the redness is measured by photography.
  • the composition increases skin hydration in a skin of an individual after applying the composition.
  • the composition promotes HA synthesis.
  • composition upregulates hyaluronic acid synthase (HAS) expression. In some embodiments, the composition upregulates hyaluronic acid synthase (HAS) expression, wherein the HAS comprises HAS2. In some embodiments, the composition downregulates hyaluronidase expression. In some embodiments, the composition downregulates hyaluronidase expression, wherein the hyaluronidase comprises hyaluronidase 2 (HYAL2).
  • the composition reduces the appearance of fine lines. In some embodiments, the composition reduces the appearance of deep lines. In some embodiments, the composition reduces the appearance of deep lines in facial skin. In some embodiments, the composition reduces the appearance of wrinkles. In some embodiments, the composition reduces the appearance of an age spot. In some embodiments, the composition reduces the appearance of crepiness. In some embodiments, the composition reduces the appearance of total area of pore size. In some embodiments, the composition reduces skin dryness. In some embodiments, the composition improves the appearance of skin texture. In some embodiments, the composition improves the appearance of solar elastosis. In some embodiments, the composition reduces or reverses a sign of solar elastosis.
  • the compositions provided herein act to increase production of high MW HA in the skin. In some embodiments, the compositions provided herein act to increase the amount of high MW HA in the skin. In some embodiments, the compositions provided herein act to increase production of high MW HA by one or more of epidermal cells, fibroblasts, keratinocytes, or other skin cells. In some embodiments, the compositions provided herein act to increase deposition of high MW HA in the skin ECM. In some embodiments, application of the compositions provided herein result in an increased ratio of high MW HA to low MW HA in the skin.
  • application of the compositions provided herein result in an increased ratio of high MW HA to low MW HA in the epidermis, dermis, or a combination thereof.
  • the increased ratio of high MW HA to low MW HA in the skin result in reduced inflammation.
  • the increased ratio of high MW HA to low MW HA in the skin result in reduced visible redness.
  • the increased ratio of high MW HA to low MW HA in the skin result in reduced appearance of visible fine lines.
  • the increased ratio of high MW HA to low MW HA in the skin result in reduced appearance of visible deep lines.
  • the increased ratio of high MW HA to low MW HA in the skin result in reduced appearance of visible deep lines in the face of an individual. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in reduced skin crepiness. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in improved skin elasticity. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in reduced total area of pore size. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in improved visible skin texture. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in reduced skin dryness.
  • the increased ratio of high MW HA to low MW HA in the skin result in increased skin hydration. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in reduced visible sign of solar elastosis. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in reversal of visible sign of solar elastosis. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in remodeling of dermal ECM to produced thicker collagen fibers. In some embodiments, the increased ratio of high MW HA to low MW HA in the skin result in remodeling of dermal ECM to produce denser collagen fibers.
  • the increased ratio of high MW HA to low MW HA in the skin result in remodeling of dermal ECM into a healthier collagen fiber network.
  • the increase in high MW HA in the skin results in a decrease in inflammation in the epidermis and an increase in high MW HA production in the dermis.
  • Stability testing of the compositions can be conducted as follows. High temperature testing is now commonly used as a predictor of long-term stability. High temperature testing can be conducted at 37°C (98°F) and 45°C (113°F). If a product is stored at 45 °C for three months (and exhibits acceptable stability) then it should be stable at room temperature for two years. A good control temperature is 4°C (39°F) where most products will exhibit excellent stability. Sometime, the product is subjected to -10°C (14°F) for three months.
  • stability of the product is assessed by passing three cycles of temperature testing from -10°C (14°F) to 25°C (77°F). In such cases, the product is placed at - 10°C for 24 hours and then placed at room temperature (25°C) for 24 hours. This completes one cycle. An even more rigorous test is a -10°C to 45°C five-cycle test. This puts emulsions under a tremendous stress.
  • the dispersed phase (of an oil-in-water emulsion) may separate and rise to the top of the emulsion forming a layer of oil droplets. This phenomenon is called creaming.
  • Creaming is one of the first signs of impending emulsion instability.
  • one test method to predict creaming is centrifugation.
  • the emulsion is heated to 50°C (122°F) and centrifuged for thirty minutes at 3000 rpm.
  • the emulsion is inspected for signs of creaming.
  • formulas and packaging can be sensitive to the UV radiation.
  • the product is placed in glass and the actual package in a light box that has a broad-spectrum output.
  • another glass jar completely covered with aluminum foil serves as a control.
  • a discoloration of the product may be observed.
  • Described herein are methods of preparing and using a composition comprising one or more peptides comprising an amino acid sequence feature found in collagen and/or elastin.
  • the peptide comprises the amino acid sequence features found in elastin.
  • the peptide comprises the amino acid sequence features found in collagen.
  • methods comprise topical application of the composition.
  • the one or more peptides comprise an octapeptide.
  • the octapeptide comprises an amino acid sequence H - Gly - Xi - X2 - X3 - X4 - X5 - Xe - X7 - OH, wherein Xi is Pro, Asp, or Leu; wherein X2 is His, Gly, Met, or Gin; wherein X3 is Gly, Asp, Pro, or Lys; wherein X4 is Vai, Gly, Pro, Phe, or Asn; wherein X5 is Arg, Ala, Ser, Gin, or Gly; wherein Xe is Glu, Ser, Gly, Arg, or Asp; and wherein X7 is Ala, Pro, or Asp.
  • the octapeptide comprises an amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD. In some embodiments, the octapeptide comprises an amino acid sequence GPHGVREA, GDGDGASA, GPMGPSGP, GLGPGARA, GPQGFQGP, GPMGPRGP, or GPGKNGDD, or any combination thereof. In some embodiments, the octapeptide comprises an amino acid sequence GPHGVREA. In some embodiments, the octapeptide comprises a chemical structure of octapeptide-45 (FIG.
  • Described herein are methods for reducing inflammation in a skin of an individual comprising topical application of one or more peptides with amino acid sequence features found in collagen or elastin.
  • methods for reducing inflammation in a skin of an individual comprise applying a topical composition comprising an octapeptide.
  • the octapeptide comprises at least one of an amino acid sequence GDGDGASA (SEQ ID NO: 1), GPMGPSGP (SEQ ID NO: 2), GLGPGARA (SEQ ID NO: 3), GPQGFQGP (SEQ ID NO: 4), GPHGVREA (SEQ ID NO: 5), GPMGPRGP (SEQ ID NO: 6), GPGKNGDD (SEQ ID NO: 7), or GPMGPRGP (SEQ ID NO: 8).
  • the octapeptide comprises an amino acid sequence GPHGVREA (SEQ ID NO: 5).
  • the octapeptide comprises octapeptide-45.
  • the composition comprises a synthetic tripeptide.
  • the synthetic tripeptide is tetradecyl aminobutyroylvalylaminobutyric urea trifluoroacetate.
  • the composition comprises a hexapeptide.
  • the hexapeptide is hexapeptide-11.
  • the hexapeptide-11 is encapsulated in a liposome.
  • the composition comprises lactoferrin.
  • the lactoferrin is encapsulated in a liposome.
  • the composition comprises phosphatidylserine.
  • the composition comprises Tremella fuciformis extract. In some embodiments, the composition comprises hydroxymethoxyphenyl decanone. In some embodiments, the composition comprises a synthetic tripeptide, an octapeptide, a hexapeptide, lactoferrin, phosphatidylserine, Tremella fuciformis extract, or hydroxymethoxyphenyl decanone, or combinations thereof. In some embodiments, the composition is aqueous.
  • the reduction in inflammation is assessed by a reduction or a lack of increase in an expression level of nitric oxide synthase 2 (NOS2), tumor necrosis factor (TNF), interleukin 12 (IL- 12b), or cluster of differentiation-80 (CD80), or combinations thereof.
  • NOS2 nitric oxide synthase 2
  • TNF tumor necrosis factor
  • IL- 12b interleukin 12
  • CD80 cluster of differentiation-80
  • the composition increases CD44 expression.
  • the composition reduces redness of a skin of an individual after applying the composition. In some embodiments, the redness is measured by photography.
  • the present disclosure relates to methods or treating or restoring aging skin or increasing elastin production in the skin, such methods comprising administering a topical composition disclosed herein to the skin.
  • the composition comprises tripeptide-1, hexapeptide- 12, and octapeptide-45.
  • the composition increases elastin production.
  • the composition increases production of tropoelastin.
  • the composition increases production of microfibrillar-associated protein 4 (MFAP4).
  • MFAP4 microfibrillar-associated protein 4
  • the composition upregulates SAI 002 expression.
  • the composition upregulates LAMA3 expression.
  • the composition upregulates MERTK expression.
  • the composition upregulates ITGB4 expression.
  • the composition upregulates gene expression for one or more pathways related to ECM regulation.
  • a composition increases skin hydration in a skin of an individual after applying the composition.
  • the skin hydration in the skin of an individual is measured at baseline.
  • the skin hydration in the skin of an individual is measured at Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 after initiation of a skin treatment routine comprising application of a topical composition.
  • an individual completes a self-assessment of skin hydration at baseline.
  • an individual completes a self-assessment of skin hydration at Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 after initiation of a skin treatment routine comprising application of a topical composition.
  • an investigator completes an assessment of skin hydration of an individual at baseline. In some embodiments, an investigator completes an assessment of skin hydration of an individual at Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 after initiation of a skin treatment routine comprising application of a topical composition. In some embodiments, an individual has a measurement taken on a side of the face using a skin hydration sensor measurement system by Wearifi. In some embodiments, an individual has a measurement taken at baseline. In some embodiments, an individual has a measurement taken at Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 after initiation of a skin treatment routine comprising application of a topical composition.
  • an individual has a skin hydration measurement taken 15 minutes post cleansing the facial skin of the individual. In some embodiments, an individual has a skin hydration measurement taken following a first 15 minutes post cleansing the facial skin of the individual and a second 15 minutes post application of a composition. In some embodiments, methods are described wherein an individual has a skin hydration measurement taken three times on a side of the face. In some embodiments, an individual has a skin hydration measurement taken three times on a side of the face in the same facial area. In some embodiments, an individual has a skin hydration measurement taken on the right and left sides of the face. In some embodiments, an individual has a skin hydration measurement taken on the right and left sides of the face, three times on each side.
  • an individual has a skin hydration measurement taken on the right and left sides of the face, three times on each side in the same facial area. In some embodiments, an individual has a skin hydration measurement that captures the water content across stratum corneum and upper epidermis. In some embodiments, an individual has a skin hydration measurement wherein the measurement is a quantitative result of volumetric ratio of water in skin tissue. In some embodiments, an individual has a skin hydration measurement wherein the measurement represents an absolute value with direct relevance to the effective hydration of the skin. In some embodiments, an individual has a skin hydration measurement taken at baseline and after initiation of a skin treatment routine comprising application of a topical composition.
  • an individual completes a self-assessment survey on features of their facial skin.
  • the survey is completed at baseline, at Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the survey is completed post-cleansing the skin.
  • the survey is completed about 15 minutes post-cleansing the skin.
  • the individual completes the survey after initiation of a skin treatment routine comprising application of a topical composition.
  • the individual completes the survey using a scale to rate the assessment of their skin.
  • the individual completes the survey using a 5-point scale to rate the assessment of their skin.
  • the individual completes a self-assessment survey on features of their facial skin at baseline and after initiation of a skin treatment routine comprising application of the composition.
  • an individual completes a self-assessment survey on overall satisfaction with the composition. In some embodiments, an individual completes a selfassessment survey on overall satisfaction with the composition. In some embodiments, an individual completes a self-assessment survey on overall satisfaction with the composition at baseline and at other time points. In some embodiments, an individual completes a selfassessment survey on overall satisfaction with the composition at baseline and Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, an overall satisfaction survey uses a scale compared to baseline. In some embodiments, an overall satisfaction survey uses a 7-point scale compared to baseline.
  • an investigator completes an assessment on the facial skin of an individual.
  • the individual initiates a skin treatment routine comprising application of the composition provided herein.
  • methods are described wherein an investigator completes an assessment on the facial skin of an individual at baseline and a later time point.
  • the later time point is Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 after the first application.
  • an investigator completes an assessment on the facial skin of an individual post cleansing of the skin.
  • an investigator completes an assessment on the facial skin of an individual about 15 minutes post cleansing of the skin.
  • an investigator completes an assessment on the facial skin of an individual to assess fine lines/wrinkling, crepiness, texture, erythema, dryness or moisture/hydration. In some embodiments, an investigator completes an assessment on the facial skin of an individual to assess fine lines/wrinkling, crepiness, texture, erythema, dryness and moisture/hydration. In some embodiments, a scale is used to assess fine lines/wrinkling, crepiness, texture, erythema, dryness and/or moisture/hydration. In some embodiments, a 10- point scale is used to assess fine lines/wrinkling, crepiness, texture, erythema, dryness and/or moisture/hydration. In some embodiments, the scale parameters comprise: 0 absent; 1-3 mild, 4-6 moderate, 7-9 severe. In some embodiments, methods are described wherein the results of the investigator assessment are subjected to statistical analysis.
  • a photograph is taken of an individual who has initiated a skin treatment routine comprising application of the composition provided herein.
  • photographs are taken showing the facial skin of an individual at baseline and at Week 2, Week 4, and/or Week 8 following initiation of a skin treatment routine comprising application of the composition.
  • a portrait lens 85mm f/1.4
  • a photograph is taken post-cleansing of the skin.
  • a photograph is taken about 15 minutes post-cleansing of the skin.
  • a photograph is taken post-cleansing of the skin and post-application of a topical study composition. In some embodiments, a photograph is taken following a first time period of about 15 minutes post-cleansing of the skin and followed by a second time period of about 15 minutes post-application of a topical study composition.
  • a VISIA® Skin Analysis System is used to take photographs. In some embodiments, a VISIA® Skin Analysis System is used to conduct analysis of skin features on photographs taken. In some embodiments, a LifeViz® Infinity System is used to take photographs. In some embodiments, a LifeViz® Infinity System is used to conduct analysis of skin features on photographs taken.
  • a LifeViz® Micro System is used to take photographs. In some embodiments, LifeViz® Micro System is used to conduct analysis of skin features on photographs taken. In some embodiments, skin redness visible in photographs of an individual is assessed. In some embodiments, skin inflammation visible in photographs of an individual is assessed. In some embodiments, skin wrinkles visible in photographs of an individual are assessed. In some embodiments, fine lines visible in photographs of an individual are assessed. In some embodiments, deep lines visible in photographs of an individual are assessed. In some embodiments, deep lines in facial skin visible in photographs of an individual are assessed. In some embodiments, crepiness in facial skin visible in photographs of an individual is assessed. In some embodiments, texture of facial skin visible in photographs of an individual is assessed.
  • pore size area of facial skin visible in photographs of an individual is assessed.
  • rosacea of facial skin visible in photographs of an individual is assessed.
  • fine blood vessel of facial skin visible in photographs of an individual are assessed.
  • solar elastosis of facial skin visible in photographs of an individual is assessed.
  • an age spot on facial skin visible in photographs of an individual is assessed.
  • a punch biopsy may be taken periauricularly at baseline, and post application at week 2, week 4 and week 8.
  • an individual may be instructed to apply a topical serum periauricular twice a day for the duration of the study.
  • an independent dermatopathologist evaluates the tissue pre and post application of a study product.
  • methods are described wherein biopsy samples are processed for histological analysis.
  • the histological analysis comprises CD44 immunohistochemistry, hemotoxylin and eosin staining.
  • hemotoxylin and eosin staining reveal the structure of dermal extracellular matrix and a thickness and density of collagen fibers.
  • CD44 immunohistochemistry reveal a cellular location of HA production.
  • the topical application of composition reduces inflammation in an individual. In some embodiments, methods are described wherein the topical application of the composition reduces skin inflammation in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the topical application of composition reduces skin redness in an individual. In some embodiments, methods are described wherein the topical application of the composition reduces skin redness in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the topical application of composition reduces the appearance of deep lines in an individual.
  • methods are described wherein the topical application of the composition reduces the appearance of deep lines in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the topical application of composition reduces the appearance of wrinkles in an individual. In some embodiments, methods are described wherein the topical application of the composition reduces the appearance of wrinkles in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the topical application of composition reduces the appearance of crepiness in an individual. In some embodiments, methods are described wherein the topical application of the composition reduces the appearance of crepiness in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the topical application of composition reduces skin dryness in an individual. In some embodiments, methods are described wherein the topical application of the composition reduces the skin dryness in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the topical application of composition increases skin hydration in an individual. In some embodiments, methods are described wherein the topical application of the composition increases skin hydration in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the topical application of composition improves the appearance of skin texture in an individual. In some embodiments, methods are described wherein the topical application of the composition improves the appearance of skin texture in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the topical application of composition improves the appearance of solar elastosis in an individual.
  • methods are described wherein the topical application of the composition improves the appearance of solar elastosis in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the topical application of composition reverses the appearance of solar elastosis in an individual.
  • methods are described wherein the topical application of the composition reverses the appearance of solar elastosis in an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the topical application of composition stimulates HA synthesis in the skin of an individual. In some embodiments, methods are described wherein the topical application of the composition stimulates HA synthesis in the skin of an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the topical application of composition upregulates hyaluronic acid synthase (HAS) expression in the skin of an individual. In some embodiments, methods are described wherein the topical application of the composition upregulates hyaluronic acid synthase (HAS) expression in the skin of an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • HAS hyaluronic acid synthase
  • the stimulation of hyaluronic acid synthase (HAS) activity in a skin of an individual comprises an increase in expression of HAS2.
  • the topical application of the composition downregulates hyaluronidase expression in the skin of an individual.
  • methods are described wherein the topical application of the composition downregulates hyaluronidase expression in the skin of an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the composition downregulates hyaluronidase expression in a skin of an individual comprises a decrease in expression of HYAL2.
  • the topical application of the composition reduces appearance of a bruise, an aging spot, or a wrinkle in a skin of an individual.
  • methods are described wherein the topical application of the composition reduces appearance of a bruise, an aging spot, or a wrinkle in a skin of an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the reduction in appearance of a bruise, an aging spot, or a wrinkle persists after the period of topical application of the composition has ended.
  • the topical application of composition increases elastin production in the skin of an individual.
  • methods are described wherein the topical application of the composition stimulates elastin production in the skin of an individual when comparing baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the topical application of the composition upregulates MF AIM expression.
  • the topical application of the composition increases tropoelastin production.
  • methods are described wherein the topical application of the composition upregulates MF AIM expression and/or tropoelastin production in the skin of an individual when compared to baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the composition upregulates expression of S100A2, LAMA3, MERTK, or ITGB4.
  • methods are described wherein the topical application of the composition upregulates expression of S100A2, LAMA3, MERTK, or ITGB4 in the skin of an individual when compared to baseline or another time point to Week 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
  • the increase in elastin production, increase in tropoelastin production or upregulation of MFAP4, S100A2, LAMA3, MERTK, or ITGB4 persists after the period of topical application of the composition has ended.
  • topical application of composition is applied 1, 2, 3, 4, 5, 6, 7, or 8 times a day.
  • the topical application is applied in the AM.
  • the topical application is applied in the PM.
  • the topical application is applied in the AM and PM.
  • the topical application is applied in the AM or PM.
  • the topical application is applied post-cleansing of the skin.
  • the topical application is applied a certain time period after post-cleansing of the skin. In some embodiments, the topical application is applied about 15 minutes post-cleansing of the skin.
  • the present disclosure relates to a method for promoting wound healing, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for promoting wound healing or wound closure, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for treating a cutaneous wound, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the wound (e.g., cutaneous wound) is related to use of a device used for anti-aging indications.
  • the anti-aging indications may include combatting skin laxity, combatting diminished skin texture and tone, reducing redness, reducing skin blemishes, reducing inflammation, reducing swelling, reducing itching of skin, or any combination thereof.
  • the present disclosure relates to a method for promoting elastogenesis, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing scarring, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing the effects of aging on skin, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing or reversing aging of skin, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing skin laxity, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for improving skin texture or improving skin tone, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for improving pigmentation, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing redness of skin, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing skin blemishes or improving the appearance of skin blemishes, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing inflammation of skin, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing swelling of skin, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for reducing itching of skin, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • the present disclosure relates to a method for promoting extracellular matrix (ECM) remodeling, comprising administering the topical composition according to the present disclosure to the skin of a subject.
  • ECM extracellular matrix
  • the skin affected by itching is not suffering from one or more of: a wound, skin damage, signs of aging, bruising (e.g., suffering from a bruise related to a body procedure such as injection of a filler), inflammation, redness, soreness, sensitivity, dryness, laxity, crepiness, poor body contouring, discoloration, swelling, a pigmentation disorder (e.g., hyperpigmentation), age spots, wrinkles, pain, discomfort, or dermatoporosis.
  • bruising e.g., suffering from a bruise related to a body procedure such as injection of a filler
  • inflammation redness, soreness, sensitivity, dryness, laxity, crepiness, poor body contouring, discoloration, swelling
  • a pigmentation disorder e.g., hyperpigmentation
  • age spots wrinkles, pain, discomfort, or dermatoporosis.
  • the individual receiving the topical application is a human.
  • kits comprising the composition comprising the peptides described herein.
  • kits can be provided to an administering physician, other health care professional, a patient, or a caregiver.
  • a kit comprises a container which contains the compositions in a suitable topical composition, and instructions for administering the composition to a subject.
  • the kit can contain one or more additional therapeutic or other agents.
  • a kit containing a composition in topical form can be provided along with other skin care agents, such as, cleansers, occlusive moisturizers, penetrating moisturizers, sunscreens, sunblocks, and the like.
  • the kit may comprise the composition in bulk form or may comprise separate doses of the composition for serial or sequential administration.
  • the kit can optionally contain one or more diagnostic tools, administration tools, and/or instructions for use.
  • the kit can contain suitable delivery devices, such as, syringes, pump dispensers, single dose packets, and the like, along with instructions for administering the peptide compositions and any other therapeutic or beneficial agents.
  • the kit can optionally contain instructions for storage, reconstitution (if applicable), and administration of any or all therapeutic or beneficial agents included.
  • the kits can include a plurality of containers reflecting the number of administrations to be given to a subject, or the different products to be administered to the subject.
  • the composition stimulates the skin regenerating process and assists in improving the skin’s appearance, and skin tightness.
  • the composition is suitable for all skin types and post-procedure skin.
  • the composition may be provided to the patient in bulk form, to permit a suitable amount of the peptides to be self-administered by the patient.
  • the patient can apply an amount of the composition sufficient to provide an even coating over the affected area or as otherwise instructed by the physician.
  • a cleanser, a sunblock, a sunscreen, a penetrating moisturizer, and/or an occlusive moisturizer can be provided for administration before or after the topical composition of the embodiments.
  • creams, ointments, lotions, solutions, gels, sprays and patches may incorporate the compositions as described herein, in combination with penetration enhancing agents and other agents to improve skin health.
  • each of the expressions “at least one of A, B and C”, “at least one of A, B, or C”, “one or more of A, B, and C”, “one or more of A, B, or C” and “A, B, and/or C” means A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B and C together.
  • “or” may refer to “and”, “or,” or “and/or” and may be used both exclusively and inclusively.
  • the term “A or B” may refer to “A or B”, “A but not B”, “B but not A”, and “A and B”. In some cases, context may dictate a particular meaning.
  • the terms “increased”, “increasing”, or “increase” are used herein to generally mean an increase by a statically significant amount.
  • the terms “increased,” or “increase,” mean an increase of at least 10% as compared to a reference level, for example an increase of at least about 10%, at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, standard, or control.
  • Other examples of “increase” include an increase of at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least 100-fold, at least 1000-fold or more as compared to a reference level.
  • “decreased”, “decreasing”, or “decrease” are used herein generally to mean a decrease by a statistically significant amount.
  • “decreased” or “decrease” means a reduction by at least 10% as compared to a reference level, for example a decrease by at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% decrease (e.g., absent level or non-detectable level as compared to a reference level), or any decrease between 10-100% as compared to a reference level.
  • a marker or symptom by these terms is meant a statistically significant decrease in such level.
  • the decrease can be, for example, at least 10%, at least 20%, at least 30%, at least 40% or more, and is preferably down to a level accepted as within the range of normal for an individual without a given disease.
  • the cells were exposed to the following compounds for 24 hr: Lactoferrin (500 pg/ml), TCVRRAF (100 pg/ml) (amino acid sequence extracted from Lactoferrin), Tri-peptide-1 - lOOppm (2.9 pg/ml), Hexapeptide 12, lOOppm (2.9 pg/ml) , TriHex combination 200ppm (2.9 pg/ml each), Hexapeptide 11 (100 pg /ml), Tranexamic acid 5% (500 pg/ml), Octapeptide (100 pg/ml) - proprietary peptide designed by R&D at Alastin (Alastin Skincare, Inc., a Galderma Company, Carlsbad CA), Phosphatidylserine 500 pg/ml, or Cannabidiol (CBD) 100 pg/ml.
  • the control cells were left untreated.
  • RNA Lysate preparation After 24 hours of compound exposure, the media was removed, the cells were washed IX with PBS. lOOul of RNA Lysis Buffer (Takara Bio Cat Num 635013, “10X RNA lysis buffer”, diluted to IX) was added to the well and mixed thoroughly by trituration, combined in RNAse free microcentrifuge tubes and immediately frozen at -30C. Samples were prepared from one cell line (one plate) at a time. Plate array is 12 x 4, with each treatment in a row of 3 wells. The triplicate wells were lysed and combined into a tube in a PCR tube strip. All samples were shipped frozen on dry ice for RNA extraction, library construction and sequencing to 25M paired end lOObp reads per sample.
  • RNA-Seq All the RNA samples were shipped frozen on dry ice for RNA extraction, library construction and sequencing to 25M paired end lOObp reads per sample. Differentially expressed genes were identified and pathway enrichment was assessed using the Reactome Pathway.
  • FIG. 1 shows data of Hyaluronic Acid Synthase 2 (HAS2), which is a primary stimulant of HA in fibroblasts, in fibroblasts treated with the various compounds.
  • HAS2 Hyaluronic Acid Synthase 2
  • FIG. 1 shows data of Hyaluronic Acid Synthase 2 (HAS2), which is a primary stimulant of HA in fibroblasts, in fibroblasts treated with the various compounds.
  • the data are presented as the fold-change in gene expression relative to the non-treated cells.
  • Octapeptide showed excellent stimulation of Hyaluronic Acid Synthase 2 (HAS2) (FIG. 1).
  • FIG. 2A shows data of HAS2 in keratinocytes treated with the various compounds. The data are presented as the fold-change in gene expression relative to the non-treated cells.
  • FIG. 2B shows data of Hyaluronidase 2 (HYAL2), which is a HA reducing enzyme, in keratinocytes treated with the various compounds.
  • HYAL2 Hyaluronidase 2
  • Hexapeptide- 11 showed excellent upregulation of HAS2 in keratinocytes and potent downregulation of HYAL2 in keratinocytes (FIG. 2A-B).
  • Octapeptide showed modest downregulation of HYAL2 in keratinocytes.
  • the data are presented as the fold-change in gene expression relative to the non-treated cells.
  • FIG. 3 shows data of Early Growth Response 3 (EGR3), in keratinocytes treated with the various compounds.
  • EGR3 is a late epidermal regulator of differentiation highly expressed in the stratum granulosum that regulates expression of skin barrier genes and can function to strengthen the barrier of the skin. The data are presented as the fold-change in gene expression relative to the non-treated cells.
  • This Example shows that the peptides described herein are involved in regulating gene expression of genes involved in hyaluronic acid stimulation, hyaluronic acid turnover and maintenance, integrity and strength of the epidermal skin barrier.
  • Example 2 Selection of Agents for HA Production and Size Assessment In Vitro
  • octapeptide, SymDecanoxTM, Tremella, lactoferrin, phosphatidylserine, HylasomeTM, AquaxylTM and full formulation stimulate the secretion of high-molecular weight (high MW) hyaluronic acid (HA) from dermal fibroblasts (and keratinocytes).
  • high MW high-molecular weight
  • HA hyaluronic acid
  • assays for HA synthesis Provided herein are assays for HA synthesis.
  • octapeptide (proprietary peptide) upregulates Hyaluronic Acid Synthase-2 (HAS2) gene expression in fibroblasts (based on RNA-Seq data described in Example 1).
  • Lactoferrin may provide wound healing attributes, promotes fibroblast proliferation and increases HA secretion.
  • Syn-Hycan is a synthetic tripeptide that stimulates HA. Published evidence suggests that a synthetic tripeptide tetradecyl aminobutyroylvalylaminobutyric urea trifluoroacetate restores facial skin volume by stimulating HA synthesis. Syn-Hycan increases HA and CD44 in the skin in vitro and ex vivo.
  • Phosphatidylserine promotes HA synthesis.
  • Hyaluronate Crosspolymer is an extremely high MW synthetic HA with exceptionally high water-binding capacity resulting in excellent moisturizing abilities.
  • Tremella fuciformis extract derived from an edible mushroom and may provide high levels of moisture from as a natural HA stimulant and has antioxidant properties.
  • Hydroxymethoxyphenyl decanone is an HA booster, antioxidant, and anti-irritant. It may stimulate the dermal and epidermal hyaluronic acid levels by 259% and 198% versus placebo, respectively, in an ex vivo human skin model.
  • Hexapeptide- 11 may upregulate HAS2 and downregulate the HA reducing enzyme hyaluronidase 2 (HYAL2) in keratinocytes (based on RNA-seq data in Example 1).
  • Table 1 Compounds Used to Treat Fibroblasts for HA Production Assessment by PAGE
  • Dermal fibroblasts were cultured in growth medium until near confluence in 6-well plates. When they reached confluence, the growth medium was replaced with serum-free medium for 24 hours. Then, the cells were treated with the compounds indicated in Table 1. After 72 hr, lOOpl of media was collected from each treatment condition. The media was concentrated in a SpeedVac concentrator to a final volume of ⁇ 10pl. All 1 O l from each condition, were loaded onto an acrylamide gel (NuPAGE 4-12% Bis-Tris Protein Gel, Invitrogen, Waltham, MA).
  • H2M Sodium Hyaluronate, Research Grade, (HA2M) from Lifecore Biomedical (Chaska, MN) was reconstituted as recommended by the manufacturer and served as a MW reference, representing high MW HA.
  • the gel was run at 200V for 3 hours to separate MW sizes from proteins and ECM components isolated from the supernatants along with the HA2M reference as a comparator.
  • Gel buffer was exchanged with fresh room temperature buffer every 20 minutes to avoid overheating.
  • the gel was stained using Stains- All (Millipore Sigma, Burlington, MA) and destained according to the manufacturer’s protocol. A band running at the same size of the HA2M band indicated the production of high MW HA.
  • FIG. 4 Data is seen in FIG. 4 showing the effect of several compounds on hyaluronic acid production in human dermal fibroblast cells 72 hours after treatment.
  • Number 1-10 are the name of the compounds listed above in Table 1.
  • Number 11 is the concentrated supernatant of untreated cells.
  • Number 12 is the HA2M control (2 MDa /high MW HA) shown as a reference point.
  • FIG. 4 demonstrates fibroblasts producing HA in a range at 2MDa/high MW HA and none produce low molecular weight (LMW) HA. Confirmation of HA
  • Fibroblasts were cultured as described above until near confluence in 6-well plates. Then, the cells were treated with the compounds indicated in Table 1. After 72 hr, lOOpl of media was collected from each treatment condition. The media was then subjected to hyaluronidase enzyme treatment (Img/ml) at 37°C for 2 hr. One condition was left non-digested (octapeptide at lOOpg/ml). The samples were run on an SDS-PAGE gel, stained, and destained as described above. Results are shown in FIG. 5. Lanes 1-8 correspond to treatment using sample numbers 1- 8 listed in Table 1. Lane 9 corresponds to octapeptide treatment at lOOpg/ml left undigested. Lane 10 is the HA2M control (2MDa high MW HA) shown as a reference point. The absence of the band after digestion indicated that the band identified in the prior step was HA having a high molecular weight (of about 2 MDa).
  • This example involved a study on the progressive effects of treatment using a topical HA and octapeptide formulation as part of a facial skin care routine in a third experiment.
  • the formulation tested included topical application of the composition comprising high MW HA and octapeptide, SPF 30+, an optional ultra-light moisturizer post Week 4, and regular skin care using a gentle cleanser in a third experiment.
  • This study evaluated the efficacy and safety of the composition comprising high MW HA and octapeptide in facial skin.
  • Photographic analysis and histological analysis of biopsies using the treatment formulation a topical application of the composition comprising high MW HA and octapeptide, SPF 30+, an optional ultra-light moisturizer post Week 4, also constituted an aspect of the third experiment.
  • Skin Hydration Sensor Measurements At every visit, participants had 3 measurements on the right and left sides of the face using a skin hydration sensor measurement system by Wearifi (Chicago, IL). 15 minutes post cleansing the facial skin and 15 minutes post application of the topical study product. Measurements were performed in the same area at every visit. The measurement depth of the sensor is optimized to 50 pm, capturing the water content across stratum comeum and upper epidermis. The measurement is a quantitative result of volumetric ratio of water in skin tissue, which represents an absolute value with direct relevance to the effective hydration.
  • participant Assessments and Satisfaction At all follow-up visits, post 15 minutes of cleansing the skin, participants completed an assessment of their facial skin (using a 5-point scale) and overall satisfaction with the topical study product (using a 7-point scale) compared to baseline. And at every visit, post 15 minutes of applying the topical serum, participants completed an assessment of their facial skin (using a 5-point scale).
  • Biopsies A 3 mm punch biopsy was performed periauricular at baseline, and post application at week 2, week 4 and week 8. Participants that elected to biopsies were instructed to apply the topical serum periauricular twice a day for the duration of the study. An independent dermatopathologist, evaluated the tissue pre and post application of the study product.
  • Biopsy samples were fixed and sectioned for histological analysis according to standard protocols. Sectioned biopsy samples were mounted on glass slides and stained with hematoxylin and eosin to reveal epidermal and dermal cellular structure, including the structure of the dermal ECM. Sectioned biopsy samples were mounted on glass slides and labeled for CD44 protein expression using an anti-CD44 antibody via immunohistochemistry following standard protocols. CD44 labeling is indicated on the sections in a darker brown staining.
  • FIG. 14-21 are photographs, analysis and histology sections from subjects at baseline and at various time points following initiation of treatment with the composition comprising high MW HA and octapeptide, SPF 30+, an optional ultra-light moisturizer post Week 4, and regular skin care using a gentle cleanser as part of a third experiment.
  • FIG. 14A demonstrates improved appearance and decreased redness after Week 4 in a female subject, age 39.
  • FIG. 14B depicts results of VISIA® Skin Analysis System on the subject from FIG. 17A demonstrating a quantifiable reduction in erythema.
  • FIG. 15 shows photographs taken (from left to right) at baseline, at Week 2, at Week 4, and at Week 8 following treatment to demonstrate improvements to progressive effects of skin improvement in a female subject, age 35. Visible, progressive changes to improve the appearance of fine lines, skin texture and redness are evident when comparing each time point indicating that continued treatment yields progressively enhanced results.
  • FIGS. 16A-16C demonstrate the extent of improvement in quantifiable features of appearance in the subject from FIG. 15 comparing baseline and Week 8 measurements.
  • FIG. 16A-16C demonstrate the extent of improvement in quantifiable features of appearance in the subject from FIG. 15 comparing baseline and Week 8 measurements.
  • FIG. 16B shows the results of the VISIA® Skin Analysis System to indicate a measurable reduction in red areas of the face.
  • FIG. 16C quantifies the effect of treatment using the VISIA® Skin Analysis System to lead to a reduction total extent of pore size area on the face of the subject.
  • FIGS. 16D-16E demonstrate the extent of the decreased appearance in fine lines and wrinkles after 8 weeks of treatment. Particularly prominent is the decrease in appearance of fine lines and wrinkles in skin around the eyes.
  • FIG. 16E quantifies the effect of treatment using the VISIA® Skin Analysis System to lead to a reduction the visibility of fine lines and wrinkles on the face of the subject.
  • FIGS. 17A-17B show photographs and analysis at baseline and at Week 8 following treatment to demonstrate improvements to the appearance of skin texture in a female subject, age 38.
  • FIG. 17B quantifies the effect of treatment using the VISIA® Skin Analysis System to lead to an improved appearance of skin texture on the face of the subject.
  • FIGS. 18A-18C show photographs demonstrating the effects of treatment to show a noticeable reduction in the appearance of erythema and rosacea in a male subject, age 48.
  • FIG. 18A shows the progressive effects of treatment when comparing baseline to Week 2, Week 4, and Week 8 in visible redness reduction in a male subject, age 48.
  • FIGS. 18B-18C show in the subject from FIG. 18A close-up views to illustrate the extent of visible reduction in erythema and rosacea in both the left and right cheek areas at Week 8. Particularly evident is the reduction in visibility of fine blood vessel following this treatment period.
  • FIGS. 19A-19C show the progressive effects of reducing the visibility of deep wrinkles near the mouth, fine lines throughout the face and a decrease in the appearance of skin crepiness yielding a smoother and tauter skin appearance in a female subject, age 73 when comparing baseline, Week 2, Week 4 and Week 8.
  • FIGS. 20A-20C show a histological analysis of periauricular biopsies of three subjects at baseline and at Week 8 following treatment with HA Immerse Serum (high MW HA + octapeptide).
  • FIG. 20A-20C each show hematoxylin and eosin-stained sections from biopsies from the same subject compared at baseline and at Week 8 (3 subjects analyzed in total).
  • Evident in each baseline sample is the extent of dermal solar elastosis visible as old, thin collagen fibers in the ECM.
  • the old, thin collagen fibers in the dermis have been replaced with thicker healthier collagen fibers producing a much more dense and healthier dermis and ECM.
  • FIGS. 21A-21C show as histological analysis of CD44 staining within periauricular biopsies of two subjects at baseline and at Week 8 following treatment with HA Immerse Serum (high MW HA + octapeptide).
  • FIG. 21 A shows sections from a first subject with CD44 expression labeled in brown via immunohistochemistry.
  • CD44 is a receptor for HA and has been shown to be upregulated upon increased extracellular HA
  • elevated CD44 labeling in both epidermis and dermis at Week 8 indicates increased stimulation and secretion of HA.
  • FIGS. 21B-21C show sections from a second subject with CD44 expression labeled in brown via immunohistochemistry.
  • FIG. 22 shows the chemical structure of octapeptide-45, comprising an amino acid sequence Gly-Pro-His-Gly-Val-Arg-Glu-Ala.
  • octapeptide-45 is the octapeptide included in the formulation of HA Immerse Serum.
  • compositions described herein are provides in Tables 2 and 3 of a weight concentration of a solution as expressed as % (w/w).
  • RNA lysis buffer (“10X RNA lysis buffer”, Takara Bio, Cat. Num. 635013), diluted to IX in RNAse-free microcentrifuge tubes. RNA extraction, library construction, and sequencing to 25M paired-end lOObp reads per sample were then performed. Differentially expressed genes were identified, and pathway enrichment was assessed using the Reactome Pathway database.
  • S100A2 is S100 calcium binding protein A2. It is a secreted matrisome-associated protein that is expressed in the dermis. S100A2 activates protein phosphatase 5 (PP5) in a calcium-dependent manner. PP5 is a serine/threonine phosphatase involved in oxidative stress responses. LAMA3 is laminin, alpha 3.
  • LAMA3 is the gene that encodes the alpha-3 chain of laminin 5, which is a major link between the epidermal basal cells and the papillary dermis. It initiates hemidesmosome formation and provides stable attachment of the epidermis to the dermis. Laminin 5 also accelerates the assembly of basement membranes and may enhance the recovery of damaged skin.
  • MERTK is tyrosine-protein kinase MER. MERTK tranduces signals from the extracellular matrix into the cytoplasm by binding to several ligands including LGALS3, TUB, TULP1, or GAS6.
  • MERTK plays a role in various processes such as macrophage clearance of apoptotic cells, platelet aggregation, cytoskeleton reorganization, and engulfment.
  • ITGB4 is integrin beta 4.
  • ITGB4 partnered with integrin alpha 6, is the receptor for laminin.
  • 34 integrin protein is particularly important in strengthening and stabilizing the skin because it is a component of hemidesmosomes in the epidermis.
  • Table 5 lists multiple pathways related to ECM regulation for which gene expression is upregulated by octapeptide-45 or the combination of TriHex + octapeptide-45, indicating therapeutic benefit in ECM regulation, including elastin synthesis.
  • the p-value is the result of an over-representation analysis.
  • the over-representation analysis is a statistical (hypergeometric distribution) test that determines whether certain Reactome pathways are over-represented (enriched) in the data. This test produces a probability score, which is corrected for false discovery rate using the Benjamani -Hochberg method.
  • human skin tissue was defatted and cut into a diameter of approximately 7 mm and placed in a transwell insert with cell culture media (DMEM/F12 containing 1.88 pM CaCh, 2% fetal bovine serum, 50 pM adenine, 0.02 nM triiodothyronine, 1% ITS-X (insulin, transferrin, selenium mix), 1% glutagro (a glutamine equivalent), 1% pen/strep, and 1% gentamycin). The medium was changed daily. The human skin tissue samples were equilibrated for 3 days, then treated with the compounds noted in Table 6 once daily for 7 days. After 7 days of treatment, the skin samples were fixed in formalin and embedded in paraffin following industry standard procedures. The resulting blocks were prepared and sectioned for immunofluorescent staining.
  • DMEM/F12 containing 1.88 pM CaCh, 2% fetal bovine serum, 50 pM adenine, 0.02 nM triiodothyr
  • TE tropoelastin
  • MFAP4 microfibrillar-associated protein 4
  • Microfibrillar-associated protein 4 (MF AIM) is an extracellular glycoprotein found in elastic fibers. MFAP4 actively promotes tropoelastin (TE) self-assembly and is a ligand of microfibrils and tropoelastin involved in proper elastic fiber organization. See P. Bartosz et al., 291 J. Biol. Chem. 1103-14 (Jan. 2016) ( Figure 10) (reproduced as FIG. 25). [0302] FIG. 27 is an image of immunofluorescent staining of cells with antibodies directed to MFAP4.
  • TE tropoelastin
  • FIG. 27 is an image of immunofluorescent staining of cells with antibodies directed to MFAP4.
  • MFAP4 expression increases with TriHex (second from left), octapeptide-45 (second from right), and TriHex + octapeptide-45 treatment (right).
  • the increase in MFAP4 expression with TriHex + octapeptide-45 shows an invasion of new fibrils into the papillary dermis (see also FIG. 26) that is greater than that observed for TriHex and octapeptide-45 alone.
  • the expression of MFAP4 suggests the opportunity for formation of new elastin fibers.
  • FIG. 28 is an image of immunofluorescent staining of cells with antibodies directed to TE.
  • TE expression increases with TriHex, octapeptide-45, and TriHex + octapeptide-45.
  • the increase in TE production with TriHex + octapeptide-45 shows an invasion of new elastin fibers into the papillary dermis that is greater than that observed after treatment with TriHex and octapeptide-45 alone.
  • the alignment of the fibers is more organized with the combination treatment, suggesting that the newly formed elastin fibers are more stable.

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Abstract

L'invention concerne des compositions et des méthodes comprenant un ou plusieurs peptides comprenant un octapeptide ayant des caractéristiques de séquence d'acides aminés trouvées dans le collagène et/ou l'élastine.
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